Contained use of
Genetically Modified
Organisms
The University of Reading
Safety Guide 15
The University of Reading Safety Guide 15
Contained use of Genetically Modified Organisms
Contents
1
KEY POINT PROCEDURAL SUMMARY SG15(1)
2
INTRODUCTION SG15(2)
3
DEFINITIONS SG15(2)
4
COMPLIANCE WITH THE CONTAINED USE REGULATIONS SG15(4)
4.1 Notification of premises
4.2 Establishment of a Genetic Modification Safety Committee
4.3 Risk assessment and allocation of containment level
4.4 Classification of activities
4.5 Notification of individual activities
4.6 Other regulations
5
THE SUB-COMMITTEE FOR BIOLOGICAL SAFETY SG15(5)
5.1 Constitution
5.2 Functions of SCBS
5.3 Powers of SCBS
5.4 Meetings and membership of SCBS
5.5 SCBS approval of GM work
5.6 Projects involving hazardous nucleic acids
6
RISK ASSESSMENT SG15(8)
6.1 The approved method of risk assessment
6.2 Model scheme for risk assessment of GMMs
6.3 The Brenner Scheme for hazard identification
6.4 Use of generic risk assessments
7
REGISTRATION OF WORKERS SG15(13)
7.1 GM Worker Registration Procedures
7.2 Workers on low risk projects
7.3 Workers on higher risk projects
7.3.1 Health Surveillance
7.3.2 Health Registration
8
GM PROJECT APPLICATIONS SG15(17)
8.1 Application forms
8.2 Low risk application form (GM1LR)
8.3 The standard application form (GM1STd)
8.4 Risk Assessment form GM03
8.5 Overall statement of risk and Lay Summary
8.6 Supporting information
8.7
9
Laboratory Facilities to be used for GM projects
ASSESSMENT AND APPROVALS PROCESS SG15(20)
9.1 Overview
9.2 Provisional BSO approval for projects involving low risk activities
9.3 Extensions to existing low risk projects
9.4 New or extended projects involving higher risk activities
9.5 Technical assessment groups
9.6 Notification of Higher Risk Projects
9.7 Commencing work on projects
9.8 Information, instruction and training
10 CONFIDENTIALITY ISSUES SG15(26)
10.1 Overview
10.2 Claims for Confidential Status
10.3 HSE Response
11 SUPERVISION OF STUDENTS UNDERTAKING GM WORK SG15(28)
12 REFERENCES SG15(28)
13 ACRONYMS SG15(29)
Appendix 1: Specimen project application for low risk project
Appendix 2: Specimen project application for higher risk (Class 3) project
Dr Malcolm Iosson
University Biological & Radiation Safety Officer
Extension 8887
SG15, Revised 3rd Edition, July 2005 (to take account of changes since October 2002.)
The University of Reading Safety Guide 15
Contained Use of Genetically Modified Organisms
1
KEY POINT PROCEDURAL SUMMARY
Members of the University who wish to undertake contained use work with genetically
modified organisms must comply with this Guide, which represents University Safety Policy.
The key points are:
1
Prospective Supervisors of projects involving the use of genetically modified organisms
(GMOs) (Section 3) must read this guide, and have attended one of the training sessions
(organised by the Biological Safety Officer (BSO)) on the legal requirements and
University procedures applicable to Genetic Modification (GM) work.
2
Applications to undertake GM work must be made on a University GM Project
Proposal Form (Section 8). This form must be accompanied by completed risk
assessment form.
3
The risk assessment should be supported by sufficient information (including
photocopies of important references) to allow an independent assessment of the risks.
Depending upon the overall risks of the project, the application will be assessed either
by an independent technical assessor for low risk projects, or by a technical assessment
group for higher risk projects (Section 9).
4
If approved by the independent assessor, the BSO can give temporary approval for
work to commence on low risk projects. This approval will be confirmed in writing, but
is subject to ratification at the next available meeting of the Sub-Committee for
Biological Safety (SCBS).
5
Only low risk projects are permitted for undergraduate use or taught practical classes
(Section 11/Safety Note 16)
6
Higher risk projects must be approved by SCBS, and be notified to HSE. (Section 9).
7
When work may start depends upon the class of activity (Section 9), and whether there
are any aspects of the project that are claimed as confidential (Section 10). All
Supervisors will be notified in writing when work may commence.
8
Supervisors must comply with any conditions attached to approval of the project. All
projects must be reviewed at least annually (Section 9). Revisions to the risk assessment
must be approved by SCBS.
9
All workers on GM projects must be registered with the University Health & Safety
Services office, and, for workers on higher risk projects, approved by the Occupational
Health Service before they can start work on an approved project. They must inform the
Health & Safety office when they finish work on a GM project.
10 Workers may only apply for registration for Class 3 work (ie at Containment Level 3)
after the Supervisor has certified that they are properly trained. After approval, such
workers must notify the Health & Safety Services office and the Occupational Health
Service not less than one month before the intended leaving date (Section 7/ Safety
Note 17).
11 All workers must be given sufficient information, instruction and training (including a
copy of this Guide) to allow them to work safely on the project(s) for which they are
registered. Details of training received should be kept.
Revised July 05
SG15(1)
Contained Use of Genetically Modified Organisms
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_
2
INTRODUCTION
This Guide is intended for those wishing to undertake work with genetically modified
organisms (GMOs) in contained use facilities (see Section 3 for definitions of these terms).
Such work is regulated by the Genetically Modified Organisms (Contained Use)
Regulations 2000 (henceforward referred to as The Regulations - Reference 1). These
Regulations not only implement the requirements of European Directive 98/81/EC (on the
contained use of genetically modified microorganisms), but also go further in applying
controls to the contained use of genetically modified higher organisms. The Regulations
replaced the earlier Contained Use Regulations 1992 (as amended), and came fully into force
on 15 February 2001 after a brief transitional period of overlap from the 1992 Regulations.
Users should also be aware of a complementary set of Regulations, the Genetically Modified
Organisms (Deliberate Release) Regulations (Reference 2), which apply to situations where
living GMOs are intentionally caused to enter the environment. Deliberate Release is dealt
with in Safety Guide 35 (in preparation) and various official publications. Anyone wishing to
conduct a deliberate release of GMOs to the environment should consult the BSO in the first
instance.
Further advice and interpretation of the requirements of the Contained Use Regulations is
available in the form of two Government publications that provide official guidance on the
legislation. (References 3 - 4). Copies are held by the Biological Safety Officer (extension
8887) for reference purposes.
3
DEFINITIONS
Definitions taken from the legislation are italicised within quotation marks.
•
Contained use - "an activity in which organisms are genetically modified, or in which
genetically modified organisms are cultured, stored, transported, destroyed, disposed of
or used in any other way and for which physical, chemical or biological barriers, or
any combination of such barriers, are used to limit their contact with, and to provide a
high level of protection for, humans and the environment."
•
Genetic modification - "…the alteration of the genetic material in that organism in a
way that does not occur naturally by mating or natural recombination or both …and
genetic modification occurs at least through the use of the techniques listed" (in
Schedule 2 to the Regulations).
The listed techniques include:
− "recombinant nucleic acid techniques involving the formation of new combinations of
genetic material by the insertion of nucleic acid molecules, produced by whatever means
outside an organism, into any virus, bacterial plasmid or other vector system and their
incorporation into a host organism in which they do not naturally occur, but in which
they are capable of continued propagation;"
− "techniques involving the direct introduction into an organism of heritable genetic
material prepared outside the organism, including micro-injection, macro-injection and
micro-encapsulation;"
− "cell fusion or hybridisation techniques where live cells with new combinations of
heritable genetic material are formed through the fusion of two or more cells by means
of methods that do not occur naturally."
SG15(2)
Revised July 05
Contained Use of Genetically Modified Organisms
Some similar techniques are not considered to be genetic modification activities:
− in vitro fertilisation;
− natural processes, such as conjugation, transduction or transformation, and
− polyploidy induction.
By contrast, some techniques are specifically excluded from the Regulations:
− mutagenesis
− cell fusion of prokaryotic species that can naturally exchange genetic material;
− cell fusion of cells of any eukaryotic species, including hybridomas and plant cell
fusions; and
− self-cloning, where the resulting organism is unlikely to cause disease or harm to
humans.
•
Genetically modified higher organism (GMHO) - a GMO that is not a
microorganism. (nb - this latter term is neither derived from, or used in the Regulations
or official guidance, but is used in this Guide to assist readers. The Regulations refer to
"GMOs other than microorganisms").
The overall term "GMO" thus includes both GMHOs and GMMs. Note that these groups of
organism are treated separately as far as legislation is concerned. Readers of this guide should
carefully note whether particular paragraphs concern all GMOs, only GMMs, or only
GMHOs such as plants or animals.
4
•
Genetically modified microorganism (GMM) - a microorganism that has been
genetically modified.
•
Genetically modified organism (GMO) - an organism created through use of one of
the techniques listed (defined) as "genetic modification".
•
Genetic modification work - "a contained use of a genetically modified organism."
•
Microorganism - "a microbiological entity, cellular or non-cellular, capable of
replication or of transferring genetic material, and includes a virus, a viroid and an
animal or plant cell in culture";
•
Organism - "a biological entity capable of replication or of transferring genetic
material and includes a microorganism, but does not include a human or a human
embryo."
•
Self-cloning - "…the removal of nucleic acid sequences from a cell of an organism,
which may or may not be followed by reinsertion of all or part of that nucleic acid (or a
synthetic equivalent) whether or not altered by enzymic or mechanical processes into
cells of the same species or into cells of phylogenetically closely related species which
can exchange genetic material by homologous recombination; and
self-cloning may include the use of recombinant vectors, with an extended history of safe
use in the particular organism, to manipulate and reinsert the nucleic acid sequences, but
the vectors shall not consist of any genetic elements other than those designed for vector
structure, vector replication, vector maintenance or marker genes."
COMPLIANCE WITH THE CONTAINED USE REGULATIONS
All members of the University who wish to engage in genetic modification work must comply
with the laws relating to these activities. Some of the requirements of the law apply to the
University (as a corporate body), but most apply to those in charge of (Supervisors of) genetic
modification work. The BSO has the responsibility for ensuring compliance by the University
as a corporate body and (in conjunction with the SCBS Secretary) acts as the central contact
Revised July 05
SG15(3)
Contained Use of Genetically Modified Organisms
_________________________________________________________________________________
_
point between the University and the regulatory authorities. Supervisors of GM projects have
the responsibility for ensuring compliance by all members of their project group. All GM
workers, whether they direct the work, or participate in it, must also comply with University
policy relating to genetic modification work (as set out in this Guide).
The major requirements of the Regulations are set out below.
4.1
Notification of premises (Regulation 9)
All premises where GM work is undertaken must be notified to HSE. The University was
notified as a single GM Centre under the 1992 Regulations, and this registration was
transferred when the 2000 Regulations came into force. This registration covers all existing
buildings in which GM work is undertaken. Note that any new facilities (buildings) not
previously used for GM work must be notified before they are used for the first time.
4.2
Establishment of a Genetic Modification Safety Committee (GMSC) (Regulation 16)
Every notified centre must establish a GMSC, whose sole statutory function is to advise on
risk assessments (see below). In the University, the Sub-committee for Biological Safety
(SCBS) fulfils the duties of a GMSC. (Section 5). Any changes in the composition of the
GMSC must be notified to HSE.
4.3
Risk assessment and allocation of containment level (Regulations 6 and 7)
A risk assessment must be conducted prior to undertaking any GM activity (Section 6). The
assessment must consider risks to human health, and also (for GMMs), risks to the
environment. The legislation specifies the way the risk assessment should be performed, and
the matters to be taken into account during that assessment. Once the risks have been
assessed, appropriate containment measures must be applied to control the risks. The activity
is then classified on the basis of these necessary control measures.
4.4
Classification of activities
Different criteria apply, depending upon whether the GMO is a GMM or a GMHO:
• Activities with genetically modified microorganisms (GMMs)
Activities are classified according to the level of containment required to control the risks, ie:
•
Class 1 activities require Containment Level 1;
•
Class 2 activities require Containment Level 2; and
•
Class 3 activities require Containment Level 3.
• Activities with genetically modified higher organisms (GMHOs)
These are classified on the basis of whether or not the GMHO poses "greater risks to humans
than the non-modified recipient organism(s)". This term is equivalent to the phrase "As Safe
In Containment as the non-modified parent or recipient" (ASIC), which was used in the
previous Regulations. Because of familiarity with the acronym ASIC, it has been retained in
this edition of the Guide.
4.5
Notification of individual activities (Regulations 10, 11 and 12)
All individual "higher risk" activities must be notified to HSE. (Section 9.6). Class 2, 3 (and
4) activities are regarded as "higher risk", and must therefore be notified. Where the work
involves GMHOs, only work with non-ASIC organisms must be notified. (The use of ASIC
GMHOs is regarded as low risk work.)
Class 3 (and 4) activities also require the written consent of HSE before the work can start
(Section 9.7).
SG15(4)
Revised July 05
Contained Use of Genetically Modified Organisms
4.6
Other regulations
The Regulations also include requirements for:
•
minimising exposure;
•
the use of containment measures, etc.;
•
the adoption of practices of good occupational safety and hygiene; and
•
information disclosure
Reference 1 gives an "official interpretation" plus guidance on these requirements. Please
contact the BSO (extension 8887) for more information if necessary.
5
THE SUB-COMMITTEE FOR BIOLOGICAL SAFETY (SCBS)
5.1
Constitution
The SCBS is constituted as a sub-committee of the University Health and Safety Committee.
(UHSC) (Safety Guide 2) Its sole legal duty is to function as a Genetic Modification Safety
Committee (GMSC) in compliance with Regulation 16 of the Contained Use Regulations.
The Sub-Committee also acts a source of expertise and advises on general matters of
biological safety when required.
Regulation 16 states: "A person who carries out an assessment pursuant to regulation 6 or 7
shall establish a genetic modification safety committee to advise him in relation to that
assessment for those activities." (For the purpose of this regulation, the University is regarded
as "a person").
SCBS has been mandated to oversee the health and safety aspects of all activities concerning
the use of genetically modified organisms, whether in Contained Use or Deliberate Release.
The remit of SCBS does not extend to matters such as considering activities using materials
derived from, but not containing, viable GMOs. Unless general issues of biological safety are
also involved, consideration of matters such as "GM ingredients" in food or animal feed is
therefore outside the remit of SCBS. (The COSHH Regulations apply to any potentially
hazardous materials derived from GMOs) (Safety Guide 14).
As of May 2004, SCBS may, if requested by the BSO or any member, also consider and
advise on specific matters relating to the use of non-modified biological organisms, including
consideration of proposals to undertake new work with hazardous biological agents.
5.2
Functions of SCBS
The functions of SCBS are:
•
To ensure the compliance of the University with the legislation relating to genetic
modification work;
•
to advise on risk assessments relating to genetic modification activities (the sole legal
duty under the Contained Use Regulations);
•
to advise on and (where appropriate) approve the health and safety aspects of projects
involving the use of biological agents (whether genetically modified or not) within the
University; and
•
to receive reports (and make recommendations thereon) for all relevant activities within
the University, on behalf of the University Health and Safety Committee. This includes
reports on any accidents or incidents involving GMOs or hazardous biological agents.
Revised July 05
SG15(5)
Contained Use of Genetically Modified Organisms
_________________________________________________________________________________
_
Items for consideration may be presented to the Committee by the Biological Safety
Officer or any SCBS member: see “Receiving reports”, below.
• Ensuring compliance
To ensure that (via SCBS) the University complies with the relevant legal requirements, all
proposals to undertake any “activity” (as defined in the Regulations) with genetically
modified organisms must be submitted to, and be approved by the SCBS. This requirement
applies, irrespective of whether the proposals are for contained use activities or for deliberate
release activities. SCBS also must approve any revisions to the risk assessment, which may
arise as a result of project review or the acquisition of any new information having a bearing
on the risk assessment.
• Advising on risk assessments
Advice on technical aspects of risk assessments is provided by members of the University
who are expert in the particular areas covered by the assessments. The experts may be
members of the sub-committee, or be independent assessors. The latter are deemed to be coopted members for the purpose of advising on the risk assessments. (Section 5.3).
• Approving arrangements for health and safety:
SCBS must approve the arrangements proposed for each GM activity before it may
commence at the University. SCBS approval is limited to arrangements to protect human
health and safety, and also for risks to the environment. As part of this approval process,
SCBS may make recommendations for changes in procedures and/ or local rules that would
improve the safety of the proposed project.
• Receiving reports:
SCBS may receive reports from the BSO, or from any Sub-Committee member on any aspect
of biological (GM or non-GM) work within the University. This includes reports on, and
analysis of, any accident or incident involving the use of GMOs or hazardous
microorganisms, that could have implications for human health and safety, and/or for the
safety of the environment. After receiving and considering a report, if appropriate, SCBS
would make recommendations to the University Health and Safety Committee (UHSC) for
consideration at the next meeting of that committee. In exceptional circumstances, SCBS or
the BSO would report directly to the Chairman of the UHSC, who has the power to suspend
any work within the University on health and safety grounds.
5.3
Powers of SCBS
SCBS has the power to:
•
require that changes be made to operating procedures, in order to improve the safety of
any GM project, or work involving hazardous microorganisms;
•
impose restrictions or limitations on any GM project at any time, and
•
require (via the Chairman of the UHSC, as above) that work on a project be suspended,
if evidence emerges that there is a wilful failure by an individual to comply with either
the legislation relating to GM work, or the University procedures, as set out in this
Safety Guide.
The Sub-committee also acts in a similar manner in relation to projects involving the use of
hazardous nucleic acids, such as those containing oncogenic sequences (Section 5.5), and
projects involving pathogenic microorganisms.
The remit of SCBS does not extend to other matters, such as giving final approval for work to
commence. Such a final approval may depend on HSE giving its consent for the activity to
SG15(6)
Revised July 05
Contained Use of Genetically Modified Organisms
start, or on the higher management of the University approving arrangements unconnected
with safety.
5.4
Meetings and membership of SCBS
SCBS normally meets once per term, and dates of meetings are published in the University
Calendar. The membership is comprised of the:
•
Chairman;
•
Head of Health & Safety;
•
Secretary;
•
Occupational Health Medical Adviser;
•
Biological Safety Officer (BSO); and
•
several specialist and lay members.
The specialist members are able to give advice on the technical aspects of risk assessments,
and on any problems that may arise during the course of the project. The lay members include
representatives of Schools not involved in GM work, and also of the recognised Unions. They
are able to provide an outside viewpoint, more typical of "the man in the street".
The membership of SCBS is listed in the University Calendar, and in the SCBS Contact list,
published annually by the Health & Safety Services office (also available on the Health &
Safety Services office Web Site via the route “Contacts” → “Health & Safety Committees”
→ SCBS “Current membership”.)
When independent assessors and technical experts are asked to advise on specific risk
assessments (Section 9.2), they are deemed to have been co-opted to the Sub-Committee for
this purpose. In particular, all proposals to undertake higher risk projects (Class 2 or higher)
are considered by a technical assessment group (Section 9.5).
All members of the Sub-Committee (including co-opted Assessors) have signed the
confidentiality agreement in respect of non-disclosure of information (Section 10).
5.5
SCBS approval of GM work
All project proposals must be approved by SCBS before any work may start. The assessment
and approval procedures are fully described in Section 9, but it is sufficient to note here that
the Biological Safety Officer may give provisional approval (BSO approval) to low risk
projects. Full approval of a project may only be given by SCBS. In addition, SCBS must
approve all project reviews and reassessments, which are conducted on an annual basis.
Failure to approve such a review requires that all active work on the project must stop until
such time as the review has been approved by SCBS.
• BSO approval
With the aim of minimising delay in the approval process, the Biological Safety Officer has
been empowered by SCBS to give temporary approval for work to start on suitable low risk
GM projects, ie, Class 1 for GMMs, or the equivalent for GMHOs. BSO approval is subject
to a report from an independent assessor, and must be ratified at the next meeting of SCBS
(Section 9.2) to gain full approval.
• Full approval
Revised July 05
SG15(7)
Contained Use of Genetically Modified Organisms
_________________________________________________________________________________
_
Full project approval may only be given by SCBS. Higher risk projects are normally
approved only after consideration of the report of a technical assessment group (Sections 9.4
and 9.5). However, it is important to note that work may not start on approved higherrisk projects until all necessary legal requirements have been satisfied (Section 9.6).
Once SCBS approval has been given, the only work that may be undertaken on the project is
that covered by the risk assessment, and described in the project application form. Applicants
are therefore advised to make their proposals as comprehensive as possible, to ensure that all
the planned work is covered by the initial risk assessment.
Project Supervisors should ensure that all their projects and the associated risk
assessments are reviewed at least annually. This is particularly important since data or
information obtained during the course of a project may result in one line of research being
abandoned in favour of another, resulting in a change in the overall direction of the project.
Any change in direction that is likely to take the project outside the approved risk assessment
must be subjected to a further risk assessment. Supervisors of approved projects should note
that the preparation of a new risk assessment is a legal requirement whenever the original risk
assessment ceases to be valid.
If there is a logical connection between the original project and the proposed new direction,
the Supervisor should submit the proposals as an extension to an existing project; otherwise,
the project will be considered anew. If in any doubt, Supervisors should discuss their plans
with the BSO at the earliest opportunity.
The revised assessment must be formally approved by SCBS before any new work begins. If
the project is not a low-risk project, the appropriate fee must be paid when the new activities
are notified to HSE (Section 9.6).
5.6
Projects involving hazardous nucleic acids
Some nucleic acid molecules may be oncogenic, for example those nucleic acid molecules
with expressible oncogenes. As such, they are regarded as hazardous substances within the
terms of the Control of Substances Hazardous to Health (COSHH) Regulations (Reference 3),
The law treats potentially oncogenic nucleic acids as known or suspect carcinogens under the
COSHH Regulations. The Carcinogens Approved Code of Practice would apply
(Reference 3). In addition, the nature of the materials means that health surveillance - in the
form of medical records - is appropriate, and it is therefore appropriate that such workers are
treated in the same way as “higher risk” GM workers (Section 7.1). Even though such nucleic
acids may be regarded as "suspect carcinogens", they can present similar hazards to workers
as those posed by GM work. (The hazard includes the possibility that the molecules could be
taken up by the workers’ own cells, and subsequently cause disease).
When the intention is to study hazardous nucleic acids per se without cloning them, or
incorporating them into living cells, projects involving their use are not regarded as genetic
modification. The legal controls derive from the COSHH Regulations rather than the
Contained Use Regulations, but the net effect is the same: a risk assessment must be
undertaken to ensure that exposure is prevented, or otherwise adequately controlled. Because
it is probable that most people within the University who work with such agents will also be
involved with GM work, SCBS will advise on the risk assessment for all projects involving
the use of potentially hazardous nucleic acids.
Proposals for such work must be submitted to SCBS on the standard GM1 project proposal
form, but this must make it clear that the work involves the use of potentially oncogenic
nucleic acids. If the proposals are approved, the local rules for the laboratory (Safety Guide
14) must clearly define the hazards, identify the risks, and state the control measures which
must be adopted to adequately control those risks.
SG15(8)
Revised July 05
Contained Use of Genetically Modified Organisms
Any reader wishing to work with potentially oncogenic nucleic acids should consult the
appropriate sections in the HSE/ACGM Compendium of Guidance (Reference 2):
6
•
Part 1 provides guidance on oncogenes and the COSHH Regulations;
•
Part 2B gives mores specific guidance on oncogenes; and
•
Part 3A, Annex 1, gives guidance on possible hazards and routes of transmission, with
practical advice on precautions and procedures. (The guidance also defines the term
"potentially oncogenic sequences").
RISK ASSESSMENT
In this context, "risk" means risks to human health and (for GMMs) risks to the environment.
For GMHOs, only risks to human health need be considered under the Contained Use
legislation. However, any environmental risks posed by GMHOs are covered by equivalent
Regulations made under the authority of the Environmental Protection Act. The Act and the
Regulations require that an assessment of the risks to the environment must be carried out
before an activity involving a GMHO is commenced ("activities" include acquiring, keeping,
using, storing, transport and destruction - Section 3). The assessment must take into account
whether any damage to the environment could occur - for most contained use activities, this
would be if the GMHO were to escape from containment.
6.1
The approved method of risk assessment
One of the essential requirements of the Regulations is that anyone planning to undertake an
activity involving genetic modification must assess the risks of that activity before the activity
is started. The way in which the assessment is done is now set down in the Regulations. They
specify the matters that must be taken into account (Schedule 4, Part 1); and also specify the
steps that must be taken (Schedule 5, Part II) to take those matters into account. Official
guidance on the process of risk assessment for GMMs (with worked examples) is given in
Part 2 of the ACGM Compendium of Guidance (Reference 2).
Irrespective of whether the assessment is for a GMM or a GMHO, the specified steps include:
Step 1 Identification of any harmful properties of the recipient, and, where appropriate, the
donor organism. This includes consideration of the:
− nature of any potentially harmful effects, associated either with the donor, the
recipient, the vector or the inserted genetic material;
− characteristics of the activity;
− severity of any potentially harmful effects; and
Step 2
Step 3
Step 4
Step 5
6.2
− likelihood that harmful effects will be realised.
Identification of any harmful properties associated with the vector or inserted material,
including any alteration in the recipient's existing properties.
Identification of the provisional level of risk associated with the GMOs.
Selection of containment and other protective measures on the basis of the provisional
level of risk and the characteristics of the activity.
Review and adjustment of the level of risk in the light of the above steps.
Model scheme for risk assessment and activity classification of GMMs
In line with the required steps described above, Proposers should consider the following
questions:
Revised July 05
SG15(9)
Contained Use of Genetically Modified Organisms
_________________________________________________________________________________
_
(1)
Does the work involve a recipient microorganism that is "inherently safe"?
Inherently safe microorganisms would either be non-pathogenic (ACDP Hazard Group 1), or
be disabled mutant strains of pathogens. They must lack genes for pathogenesis, and be
unable to survive in, or colonise the human body or the environment. They would commonly
have nutritional requirements that could only be supplied in special media, and would not
have characteristics (such as antibiotic resistance) that could be transferred to other
microorganisms.
Examples include:
•
non-pathogenic microorganisms such as Lactobacillus bulgaricus;
•
disabled strains of E. coli K12;
•
disarmed strains of Agrobacterium tumefaciens;
•
some mammalian cells and
•
certain strains of yeasts such as Saccharomyces and Pichia.
If the answer to question 1 is "No", detailed consideration will be required. For recipient
organisms, the ACDP hazard grouping is often sufficient to identify a provisional hazard
grouping of the GMM. Note however that the ACDP hazard grouping is based on the
likelihood and severity of human disease and/or infection. Special considerations apply to
plant pathogens, which pose risks to the environment rather than to humans: most plant
pathogens are not regarded (or listed) as biological agents - see Safety Guide 14.
Note that Plant Health legislation imposes additional controls on specified plant pathogens
(especially for non-native organisms). A licence from the Department of the Environment,
Food and Rural Affairs (DEFRA) may be required before such organisms may be imported or
used. Please contact the BSO (ext. 8887) for advice if in any doubt.
(2)
Could the modification affect any inherent hazardous properties of the recipient
organism?
The question may be interpreted as "could the insert make the recipient more hazardous, less
hazardous, or have no effect on the hazards posed"? To be able to answer this question, the
proposer needs to consider whether the insert codes for any harmful properties, or modifies
the host in a way that could increase the potential for harm. If a gene is cloned into a
microorganism, it could give rise to a harmful phenotype either by directly encoding a
harmful product (such as a toxin), or it might modify existing pathogenic properties, for
example changing the host range or tissue specificity of the GMM. It might also complement
a defective pathogenic gene in the recipient, and thereby restore a pathogenic function normal
to that organism. (This consideration is especially relevant when using a disabled mutant of a
pathogenic organism such as E. coli as an inherently safe host).
It is important to consider the question: "Could the potential for harm be greater than that
from the wild-type recipient organism?" For example, cloning and expression of the gene for
the A-subunit of the cholera toxin into a disabled strain of E. coli is unlikely to increase the
hazard to a level greater than that of wild-type E. coli or toxigenic Vibrio cholerae (both
Hazard Group 2). By contrast, expressing the A-subunit of the potent phytotoxin Ricin in
such a way that it was linked to bacterial lipopolysaccharide and capable of being presented
to cells, would probably warrant an increase to hazard group 3 - or even 4.
The properties of the vector may also need to be taken into account, if the vector (eg,
plasmid) remains within the recipient and expresses functions such as antibiotic resistance, or
could transfer to and express the inserted gene(s) in other organisms.
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If the answer to question 2 is "Yes", detailed consideration will be required.
(3)
Are there any circumstances in which the GMM would be likely to cause disease in
humans, animals or plants, or cause harm to the environment?
This stage is required if either the answer to question (1) is "No", and/or the answer to
question (2) is "Yes". The proposer should consider the worst-case scenario, ie, the worker
being accidentally inoculated with the organism, or a total loss of containment resulting in the
GMM entering the environment. (In effect, assume that no containment measures exist). Note
that disease includes the possibility of allergic responses and autoimmune disease, as well as
disease caused by infection or the direct expression of the effects of a toxin.
Consideration should also be given to the possibility that harm could be caused by transfer of
the insert to any other organism following a loss of containment. In addition to disease
causing harm to humans, animals or plants, other types of harm should be considered - for
example, could the GMM displace any natural organisms resident in the ecosystem? (This
could in itself be regarded as causing harm to the environment). Harm could also include
compromising antibiotic therapy, especially if the vector could be transferred to and then
express this resistance in other organisms.
(4)
What magnitude or severity of harm could result in these circumstances?
In respect of disease to humans, the severity depends on risks to workers and to the
community, and the availability of prophylaxis or treatment (ie, the criteria used for hazard
classification). In respect of damage to the environment (including diseases caused to
animals or plants), severity of harm must necessarily take into account the financial or other
costs imposed on persons affected, as well as damage to or loss of populations of at risk
species.
(5)
What is the likelihood that harm could result?
Estimation of likelihood is often very difficult, as there may be little quantitative data on
which to base estimates of likelihood. It may require an informed judgement to be made of
the ability of the GMM to establish an in vivo infection, or survive and spread in the
environment. It is often argued that the modifications may impose a metabolic burden on the
organism, and render it less fit than the unmodified organism. This however should not be
assumed to be the case, unless there is scientific evidence to support the claim. If the data is
poor or anecdotal, it may be necessary to adopt the worst-case scenario.
(6)
What containment measures are required to control the risks?
If the organism is inherently safe, and the insert does not increase the hazard potential of the
organism, Containment Level 1 should be sufficient. The activity can then be provisionally
allocated to Class 1. For other situations, consideration of questions (1) to (5) should allow a
provisional allocation to the Class of activity.
(7)
Could the provisional level of containment be affected by any of the planned
activities?
The provisional allocation should be reviewed in the light of the planned activities. In
particular, any non-standard operations that are not accounted for by the general requirements
for the provisional containment level must be considered. For example, the use of the GMM
at large scale, or the use of sonication equipment to break open cells may pose particular
problems that require additional control measures. At this point, Proposers should refer to
Table 1a - containment measures - in Part II of Schedule 8 to the Regulations (Reference 1) to
decide on the additional measures required.
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_
6.3
The Brenner Scheme for hazard identification
Although long-standing users of GMOs will be familiar with this scheme for "risk
assessment", the Brenner scheme is more properly a method of hazard identification, since it
does not take into account the potential for exposure to the hazard. The Brenner Scheme is no
longer regarded as a comprehensive risk assessment, and should only be used as a guide to
the level of containment required, provided that its limitations are accepted (see below).
This system uses the criteria of "Access", "Expression" and "Damage" in a numerical scheme,
with the final number being equated with a containment level:
•
Access - a measure of the ability of the host to colonise or be maintained within the
human body;
•
Expression - a measure of the level of expression of the inserted DNA; and
•
Damage - a measure of the potential for harm of the expressed protein.
Values for each criterion are given in multiples of 10-3; the greater the number, the greater the
hazard, and hence containment required. One problem is that users may be misled into
believing that it is a quantitative system, whereas it is at best qualitative, in that it only
indicates an order of magnitude for the risk.
• Limitations of the Brenner Scheme
The system was originally devised for cloning and expression work in bacteria (such as
E. coli), but is inappropriate for systems based on mammalian viral vectors. It is difficult or
impossible to apply to work where the expressed product is not a protein (for example,
ribosomal RNA), or where the insert is a deletion mutant of a gene. It should be used with
caution if, for example, pathogenic determinants are being cloned, or the work involves genes
that might exacerbate existing pathogenic traits (for example, antibiotic resistance genes that
might prejudice the clinical use of the antibiotic). The scheme does not take into account
insertional effects, whereby the recipient's properties are altered by the physical insertion of
the foreign DNA into the genome - the alteration in properties would have no relationship to
the nature of the insert. By contrast, in some cases, the containment levels indicated by the
scheme might in some cases be greatly in excess of what is warranted on the basis of other
criteria.
HSE no longer accept stand-alone use of the scheme, although they acknowledge that many
workers are familiar with it and find it useful, especially for routine cloning and expression
work in bacteria. It must be remembered that the Scheme is now only intended as a guide to
the risk assessment, and that Proposers must be able to show that they have taken into account
all the required factors, and used the required steps when undertaking the risk assessment.
The current version of the ACGM guidance (Reference 4) contains information on Access
factors etc. for many commonly used systems for cloning and expression work in prokaryotes,
but the information has not been updated since 1997. If Proposers use the system, it must
never be used as a stand-alone method of risk assessment, but only as an indicator of the
provisional level of containment. The provisional level thus identified should then be
reconsidered on the basis of whether the level is appropriate to control the risks of the
activity.
6.4
Use of generic risk assessments
HSE accept that generic risk assessments are useful for projects at the lowest risk level
(Class 1, or equivalent for GMHOs), as they allow those involved in approving risk
assessments to concentrate on the higher risk projects. HSE indicate that the majority of
routine cloning and expression projects (using well-characterised bacterial hosts and vectors)
could be treated under such generic arrangements. Provided that the inserted DNA did not
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code for any pathogenic characters (ie, characters that increased the probability that the GMM
could cause disease), a whole range of possible inserts could be incorporated without
increasing the risks (and hence containment requirements) of the proposed work.
In order to take advantage of this facility, the proposer should initially submit information on
the range of vectors hosts and inserts that could be used within the generic risk assessment
(Sections 8.1 and 9.2). If the generic project were approved, there would be no need to seek
further approval for any work that came within these agreed boundaries: the generic
assessment would be in effect be the approved project. Changes of personnel involved in the
work would however have to be approved in the normal way (Section 7).
Project Supervisors must always ensure that all low risk work being undertaken comes within
the agreed boundaries of the approved project. If it is likely to move outside those boundaries,
a new project proposal or request for an extension must be submitted. (In this situation, use of
a new vector or host would not constitute a move outside the agreed boundaries if the new
vectors/ hosts were well characterised and functionally equivalent to those already agreed,
and the risks of the activity were not affected). If in doubt, Supervisors should contact the
BSO for advice.
By contrast, higher risk projects will require a case-by-case assessment, which should
examine in detail all elements of the proposed work. The only exception to this would be
where the project incorporates some Class 1 activities, which could be treated on a generic
basis.
7
REGISTRATION OF WORKERS
7.1
GM Worker Registration Procedures
All GM workers must be both registered with the Health & Safety Services office, and (for
workers on “higher risk” [Class 2 or higher] projects, also be medically approved for work.
ALL GM work must be on an approved project. Registration includes recording project and
contact details onto the Health & Safety Services office database of GM workers. Medical
approval (if required) includes registration with the University Occupational Health Medical
Adviser (UOHMA) for health surveillance purposes. (see section 7.3)
A completed worker registration form (Form GM02) is required for each project the worker
joins. The forms are available via the Area Health & Safety Co-ordinator (HASC), or in
electronic form, via the “safety forms” page of the Health & Safety Services office Web Site:
(http://www.fmd.rdg.ac.uk/safety/forms.asp?DSID=12).
There are two versions of the registration form: GM02LowRisk, for workers on low risk
(Class 1) projects, and GM02Std, for workers on higher risk projects. Both versions request
basic contact details for the applicant, and details of the project involved (Project name,
reference number.) Details of the particular submission route to be followed depend on the
risk class of the proposed work - see below.
Once approved to commence work, the BSO will send an “approval to commence work”
letter to the applicant, and copied by e-mail to the project supervisor, the HASC and the Area
Technical Health & Safety Administrator. The HASC should retain a copy of the worker
approval letter for all GM workers in his Area. As part of the Annual Safety Audit, the HASC
is requested to inform the Health & Safety Services office (on an annual basis) of the names
of all GM workers currently registered in his/her Area.
Revised July 05
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_
7.2
Registration of workers on “Low risk” projects
• The form GM02LowRisk is used, and is completed jointly by the applicant and the project
supervisor
• The applicant is asked to supply basic contact details (including e-mail address) and a
minimum of personal details; the project supervisor is asked to supply details of the
project, and the predicted finish date for the applicant’s work on the project.
• The form is submitted to the BSO at Health & Safety Services, either by e-mail or it can be
printed and sent as hard copy – e-mail is preferred. Submission of the form equates with
the supervisor agreeing to provide any necessary training, and also agreeing to notify the
BSO of when the applicant ceases work on the project.
• On receipt, the BSO will record details in the database of GM workers, and issue an
approval letter (as above). Approval is normally given within 24 hours of receipt of the
registration request. The Approval letter will require the applicant to attend the next GM
training session organised by the BSO, unless he/ she has attended a session in the past, or
(because of a very limited period of registration), will have finished the work before the
next training session. In this situation, the responsibility for ensuring knowledge of, and
compliance with the relevant Regulations devolves to the project Supervisor. Other
conditions may also be applied to the registration.
• Approval is limited to the duration specified in the registration form, or a maximum of 5
years [Academic & technical staff/ Post-docs], 3 years [Post grads. or 1 year
[Undergrads.], whichever is the less. Renewal of registration is possible, provided that 30
days notice of renewal is given to the BSO by the project supervisor, otherwise the
registration will automatically expire at the end of the registration period.
• There are no reminders of impending expiry of registration.
7.3
Registration of workers on “higher risk” projects:
• The form GM02Std. is used, and on completion is sent directly to Occupational Health –
in “hard copy”, as the Occupational Health Service do not have the facility to handle email requests.
• If appropriate, the form must be accompanied by a completed Health Questionnaire from
the applicant (if a questionnaire has not been submitted to Occupational Health within the
last 3 years.) (see below.)
• Contact details, and project details are given, as in the “low risk” registration procedure; in
addition, the form requires the applicant to sign a declaration that he/she will inform all
recipients (see below) of any changes to the information in the form. Changes that should
be notified include:
•
change of address or contact details;
•
change of name;
•
cessation of work on the project, and
•
an intention to leave the University
• The Supervisor is also asked to sign a declaration stating that he/she will ensure that any
necessary training is provided for the worker.
• The worker registration form should be completed jointly by the applicant and by the
Project Supervisor before being copied. Each applicant should retain a copy of the form for
their own records, and send copies to the
•
Project Supervisor;
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•
HASC; and
•
UOHMA (to assist in health assessment).
If the application is approved by the UOHMA, a “letter of medical approval” is sent to the
BSO, who would issue a “permission to start work” letter to the applicant (details as
described above.)
This dual registration/ approval process is designed to ensure that all prospective workers on
higher risk GM projects are registered with, and seen by Occupational Health before they can
be issued with an “approval to commence work” letter. Approval may be delayed if a
prospective worker is requested to attend Occupational Health, but fails to attend an
appointment. In normal circumstances, “approval to commence work” can take up to 10 days
after the date of submission of the forms to Occupational Health.
7.3.1
Health Surveillance
As stated above, all “higher risk” GM workers are required to undergo health surveillance as
a condition of approval. Normally, surveillance is limited to the submission of a health
questionnaire in confidence to the UOHMA, who will assess the information in the light of
the proposed GM work. Applicants may also be required to attend for a medical interview
before being given approval.
Official guidance on health surveillance is given in Part 1 of the ACGM/HSE Compendium
of Guidance (Reference 2). The Contained Use Regulations do not impose a specific
requirement for health surveillance. However, health surveillance may be a requirement of
other Regulations, where "it is appropriate to protect the health of workers". Such regulations
include the COSHH Regulations (Reference 3), and the Management of Health and Safety at
Work Regulations (MOHSAW – Reference 4). In particular, regulation 11 of COSHH
specifically mentions health surveillance where there is a possibility of exposure to biological
agents (including GMMs).
The guidance makes clear that the circumstances in which health surveillance will be
appropriate, include situations when there are:
•
identifiable health effects that may be related to exposure;
•
valid reasons to assume that a disease or effect may occur under the conditions of the
work; and
•
valid techniques for detecting indications of the health effect.
Health surveillance may also be useful to assist in:
•
the evaluation of the measures used to control exposure;
•
the collection of data to evaluate the hazards to health; and
•
determining the immunity of workers to biological agents.
Within the University, the majority of workers will be undertaking low-risk work (Class 1, or
"ASIC"), and are not now required to be under health surveillance. However, any person who
may have either defective barriers to infection (such as eczema, skin allergies etc); is
immunocompromised as a result of steroid treatment, etc., or is in any way concerned about
the implications of the proposed work for their health is advised to contact the UOHMA for
advice.
Workers engaged in work in the highest risk class permissible at the University (Class 3) will
be required to give a baseline serum sample as well as being approved by the UOHMA before
they start work. They will also be required to attend a leaving interview and provide a leaving
Revised July 05
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_________________________________________________________________________________
_
serum sample once they (permanently) finish all Class 3 work. These requirements may also
be extended to certain Class 2 projects if appropriate.
•
Class 3 activities
Registered workers should inform the Occupational Health Service and the Health & Safety
Services office of the intention to leave or finish Class 3 work not less than 4 weeks before
the intended leaving date. This will allow the Occupational Health Service time to arrange a
leaving medical interview. The Personnel Office will not release the employment records of
Containment Level 3 workers until clearance is given by the Occupational Health Service.
See also Safety Note 17, which sets out the current practices for Containment level 3 work,
whether GM or not.
7.3.2
Health Registration procedures
To enable health surveillance to be undertaken, all “higher risk” GM workers must register
with, and be approved by, the UOHMA before commencing work on an authorised project.
(Note: if the Supervisor limits his/her involvement to direction of the activities with no
practical involvement, he/she would not be required to register as a worker on that project).
The health registration process involves each worker submitting details of his/her medical
history (in confidence) to the UOHMA by means of a Health Questionnaire. This should not
be copied or sent to anyone other than the UOHMA: the information is regarded as
confidential between the applicant and the UOHMA.
The Health Questionnaire is obtainable from the Health & Safety Co-ordinator (HASC) or the
Health & Safety Technical Adviser, depending on the local arrangements in the worker’s
Area. The completed questionnaire should be returned to the UOHMA in a sealed envelope,
together with a copy of the Worker Registration Form GM02Std. The project details on this
form assist the UOHMA to assess the risks to which the worker may be exposed, and hence
the approvals process.
It is important to note that:
8
•
the approval of the UOHMA must be received by the BSO before the worker “approval
to commence work” letter can be issued, and at least 10 days must be allowed for this
approval process;
•
the worker must inform the Health & Safety Services office and Occupational Health
when he/she finishes work on a higher risk GM project; and
•
if an individual worker finishes work on one project, and then at a later time wishes to
commence work on a different “higher risk” project, he/she must re-register with the
UOHMA and the Health & Safety Services database.
•
Re-registration as a GM worker (on the Health & Safety Services database) is still
required before work can start on a new “lower risk” project, but medical approval is
not required.
GM PROJECT APPLICATIONS
All project proposals must comply with the requirements of the relevant Regulations. Readers
are urged to contact either their Health & Safety Co-ordinator and/or the BSO (extension
8887) to check on what is currently required before submitting a formal proposal to SCBS.
Intending applicants are also recommended to consult the BSO informally to discuss any
implications of their proposals, and to ensure that they have considered all relevant factors in
both the human and environmental risk assessments.
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8.1
Application forms
All persons who wish to undertake GM work must submit an application to SCBS via the
Health & Safety Services office. Applications must be made on the appropriate Project
Proposal Form, GM1STd or GM1LR (see below), together with a completed Risk
Assessment form (GM03). Printed versions of the forms are obtainable from the Health &
Safety Co-ordinator (HASC), or the Health & Safety Technical Adviser (HASTA) of relevant
Schools. Applicants are however encouraged to use the electronic version of the form,
available via the “A to Z” of the Health & Safety Services office Web Site
(http://www.fmd.rdg.ac.uk/safety/forms.asp?DSID=12) where the latest version of each form
may be obtained. (Section 8.2) Forms can then be completed electronically, and submitted as
an e-mail attachment in the first instance. On receipt of the proposal form in the Health &
Safety Services office, the BSO will undertake an initial assessment, and decide the route for
assessment and approval. He may, if the proposal is judged unsatisfactory and unlikely to be
approved in its current form, refer it back to the proposer in order to supply more information,
or even request that the proposer completely revise the proposals before resubmitting them.
The BSO will apply a reference number, and return the attachment to the project proposer
with any comments on the application. In this way, a dialogue can be initiated, which will
result in a final version of the application being agreed by the proposer and the BSO.
Once agreed, the application forms should be printed; relevant declarations signed, and then
be formally submitted to the BSO (copied to the HASC and HASTA).
.
The two Contained Use Project Proposal Forms are:
•
Form GM1LR, which should only be used for low risk projects involving "inherently
safe" donors, recipients and vectors; and
•
Form GM1Std, which should be used for all other projects.
Both forms are used in conjunction with a risk assessment form (ref. GM03), which is
designed to lead applicants through the required steps of the risk assessment, and to assist
them in writing the Overall Statement of Risks and the Lay Summary of the project.
The design of each form is subject to change, particularly if any of the regulatory
requirements change, or if experience shows that HSE are paying attention to specific aspects
of the legislation. Proposers should ensure they use the latest version of each form, as
available on the Health & Safety Services office Web Site (as above).
8.2
Low risk application form GM1LR
This form is aimed to fast-track the assessment and approval process, and may only be used
where there is an intention to use inherently safe GMMs (or the equivalent for GMHOs) on
the project. This form asks the proposer to confirm the "low risk" status of the GMOs to be
used. (Supporting evidence in the form of references, and justification of statements made
may be required.) The questions require the proposer to identify the full range of possibilities
(genes, hosts, vectors, etc.) that will be used, and is linked to the risk assessment form GM03
(see below.)
If the project involves the use of organisms, vectors etc. not currently accepted as "inherently
safe", the standard application form (GM1Std) must be used, even if the project is
subsequently accepted as a low risk (Class 1 or equivalent) project. (A full risk assessment is
required in order to justify the low risk status of the project).
Revised July 05
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_
8.3
The standard application form, GM1Std
This form should be used for all projects not involving the use of inherently safe organisms
(etc). It asks the proposer to supply information on the project and to identify any potentially
hazardous properties of the parental, recipient or donor organisms, the vector and/ or the
inserted genetic material.
Form GM1Std is also designed to be compatible with the equivalent HSE project notification
form CU2 (Section 9.6), although there is no regulatory requirement to make notifications on
that form.
8.4
Risk assessment form, GM03
This form has been introduced to guide applicants through the required steps of the risk
assessment process, according to the "model scheme" given in Section 6 (above). Past
experience had shown that environmental risks were not thoroughly or rigorously assessed in
some cases. Form GM03 aims to identify both the magnitude and nature of the risks posed by
the work: once these are known or estimated, measures to control the risks can be devised and
put in place. The form replaces the sections of the previous project proposal forms that related
to risk assessment, so that duplication of information is minimised. For new project
applications, a project reference number should be obtained from the BSO before submitting
the form.
Applicants should provide enough information to allow a technical expert on the SubCommittee to check the risk assessment (and offer advice where appropriate). For example,
low-risk project applications should present enough detail to enable an assessor to check the
inherently safe status of the organisms involved. Note that special care is needed when
assessing the risks of projects that involve the use of eukaryotic viral vectors. It is unlikely
that any project involving a vector based on a Hazard Group 2 virus will be acceptable for a
generic low-risk project, as they are most unlikely to be "inherently safe". Care is also needed
where the level of containment required for the control of environmental risks is higher than
that required for control of risks to human health.
The form requires the proposer to produce a summary "Statement on the overall risks"
associated with each project, and a Lay Summary of the project, which summarises the aims,
methods, risks and control measures to be adopted for each project.
This form should also be used whenever an existing project is reviewed, to determine
whether there have been any changes that might have a significant effect on the risks of the
activity. Any such changes should be incorporated into a revised "Statement on the overall
risks" and Lay Summary (see below.)
Appendix 1 gives a worked example of a generic low risk project, and Appendix 2 gives a
worked example of a higher risk project.
Applicants should note that they have a legal duty to keep copies of the risk assessments (and
reviews thereof) for 10 years after the date of cessation of the GM activities to which the
assessment relates.
8.5
The Statement of overall risk and Lay Summary
These summary statement are normally the sole means of informing the lay members of
SCBS of the essential features of a project proposal, i.e.:
•
the aims and methods;
•
the main hazards and risks, both to humans and to the environment, and
•
the measures to be adopted to control those risks.
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Any comments made by the independent assessor will also be presented to the Sub
Committee.
To allow SCBS to comply with its legal duties to advise on GM risk assessments, it is
essential for Proposers to draft the above summaries very carefully in non-technical language
without the use of jargon. Summaries are normally circulated to SCBS members in advance
of the meeting, and if requested by members, project Proposers can be requested to attend the
meeting to answer questions.
SCBS may, at its discretion, delay approval if the summaries are not considered acceptable.
(Section 9, Assessment and approvals process.)
A reviewed or revised lay summary of each project must be submitted to SCBS annually in
time for the January meeting of each year. Failure to approve a project will result in the
particular project being declared "inactive",
8.6
Supporting information
For higher risk projects, the application should be accompanied by relevant documents, such
as copies of published papers referred to in the risk assessment, or, for example, copies of
relevant pages of commercial catalogues where a commercial vector system is to be used.
Thus, details of host strains and vectors should always be given for cloning experiments, and
an estimate made of the expected levels of expression for any expression work. If a variety of
hosts, vectors, etc, will be used, the information given should relate to the combination that
gives the highest risk, according to the recommended method of risk assessment.
Copies of the Local Rules (as revised) must also be included when a new project (or
extension) requires revision of any existing rules, or the creation of new ones. (Local Rules
should always provide information on any special handling techniques, as well as information
on hazards, risks, routine decontamination and waste control methods, etc - (Safety Guide 14,
Section 7).
Before formal submission, the application forms must be signed by the project proposer, and
countersigned as being approved by either the HASC or the Head of School. The person
countersigning the form must not be involved in the project. The risk assessment form
contains a declaration that must be signed by the project proposer.
All project proposals and/or risk assessments must be submitted to SCBS via the Health &
Safety Services office, marked "For the attention of SCBS".
8.7
Laboratory Facilities to be used for GM projects
All facilities must have been inspected and approved by the BSO prior to the proposed use. If
the project will involve the use of new facilities in an existing "notified building" ie a
laboratory not previously used for a GM project, this should be indicated on the proposal
form by checking the appropriate box. The Project Proposer will then need to make
arrangements for the BSO (extension 8887) to undertake an inspection of the facilities.
If the facilities are judged to be unsuitable in their present form, the BSO will recommend
that either they be upgraded, or that different facilities be used for the project. In the latter
case, a suitable minimum level of facilities will be recommended.
Similarly, if it were proposed that previously approved facilities are to be used in a new
project, where there would be a major change in direction from a previous project, the
facilities would need to be re-inspected for suitability. This would apply, for example, if all
work had previously been done with GMMs, and the new direction involved the
Revised July 05
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establishment and maintenance of a colony of genetically modified insects such as
Drosophila.
For work with GMMs, the required level of containment determines the Class of the
activity, but note that the classification does not apply to the use of GMHOs. (For GMHOs,
the relevant criterion is whether the GMHO poses greater risks to human health than does the
non-modified organism.) Note also that the required level and the level actually used are not
necessarily the same. For example, a Class 1 activity (one which only requires Containment
Level 1) remains as Class 1, even if it is undertaken in Containment Level 2 facilities for
reasons of convenience.
Where a new building is to be used for GM work for the first time, the University is required
to notify HSE of the additional facilities (Section 4.1).
9
ASSESSMENT AND APPROVALS PROCESS
9.1
Overview
The process for assessment and subsequent approval of project proposals depends on the type
of activities proposed in the project. Activities that are judged to be of low risk are subjected
to minimal requirements in order to comply with the law. By contrast, as the risk of the
activity increases, more information is required, both in the risk assessments and on the extent
of the measures to be applied to control those risks. Details of higher risk projects have to be
notified to HSE.
On receipt of the formal application, the appropriate procedures for assessment and approval
will be initiated, ie, the:
•
BSO approval route for low-risk projects (Section 9.2); or
•
technical sub-committee route for all higher risk proposals (Sections 9.4 and 9.5).
All applications accepted on the Low risk form GM1LR follow the BSO approval route.
For notifiable (higher risk) projects, the information on the University project application
form GM1 can be transferred directly to the equivalent notification form for HSE
(reference CU2), although there is no regulatory requirement to use this form. Proposers may
submit proposals for higher risk projects at any time, but sufficient time should be allowed for
the whole approval process to be completed before the projected date of starting work.
The approval process involves discussion of the proposals by appropriate technical specialists
(Section 9.4); approval by SCBS as a whole, and notification to HSE. Depending on
availability of suitable personnel, the whole process can take 8 weeks or more between
submission of the proposals, and formal acknowledgement by HSE.
Each application must be accompanied by a full risk assessment (Form GM03), both for
human health and safety and for the environment. The application forms must be completed
in full, otherwise consideration will be delayed whilst additional information is sought. All
new applications should be adequately supported by references to (or photocopies of)
published work. The importance of the summary statements has already been emphasised
(Section 8.5).
9.2
Provisional BSO approval for projects involving low risk activities
Low-risk activities are those involving either Class 1 activities (for GMMs) or equivalent
GMHOs. The BSO (or, in his absence, one of the two deputies) can give approval for work to
SG15(20)
05
Revised July
Contained Use of Genetically Modified Organisms
start, subject to advice from an independent assessor (see below). Work may start as soon as
University approval has been granted.
BSO approval is provisional, and is subject to confirmation by the next scheduled meeting of
the full SCBS. In such cases, the BSO will send a copy of the proposal to an independent
technical assessor, and await his/her assessment before giving approval. The independent
technical assessor would be a member of staff unconnected with the project, and judged
sufficiently familiar with the issues involved to be able to assess the project from the evidence
presented. He or she might not be a permanent member of SCBS, but if not, would be deemed
to be co-opted to SCBS for the purpose of assessing and advising on the project. (Section
5.4).
Normally, approvals by this route take about four weeks from the date of submission to date
of approval, although it may take longer, for example during the Summer Vacation, when
potential assessors may be away on leave. Two weeks are allowed for the independent
assessor to consider the proposals and respond with any comments, etc. Any queries that
arose would be referred back to the proposer, which could delay the approval. If the assessor
recommends approval, this decision will initially be communicated verbally to the proposer,
and confirmed in writing within a week by the SCBS Secretary. A copy of the approval letter
is sent to the Head of School, and the Health & Safety Co-ordinator.
• Ratification of BSO approval
All projects operating under BSO approval will be submitted to the first available meeting of
SCBS for ratification. At that meeting, the lay summary of the project will be presented to the
Committee for approval. (Section 8.4). During their consideration of the project, SCBS may
require changes to be made, or impose conditions for approval - for example, they may
require changes to the wording of the lay summary. SCBS may also decline to ratify the BSO
approval if they regard the lay summary as "not acceptable in its present form." In this
situation, they would normally require that the lay summary be resubmitted. As BSO
approval is provisional, and is limited to the time between granting of that approval and the
next available meeting of SCBS, such a referral has the consequence that work would have to
be suspended until SCBS approval has been obtained.
9.3
Extensions to existing low risk projects
Many existing low risk projects were approved before the introduction of the scheme for
"inherently safe" generic projects, and are subject to close restrictions on what is permitted.
The Supervisor must apply for a project extension for any proposals to move outside these
agreed boundaries.
To be considered as an extension to an existing project, there has to be a logical connection
between the original project and the proposed extension. Applications for extensions must
refer to the initial project proposal in the new application, and Proposers are encouraged to
convert the project to an "inherently safe" generic project where possible. Even if this is not
possible, applicants should always include a completed project proposal form (ref. GM1) and
risk assessment form (ref. GM03) for the proposed extension.
Proposals approved as low risk generic projects would not require an application for an
extension if the proposed new work were fully covered by the existing risk assessment.
If the risk assessment of a proposed extension indicates that there could be a higher risk
(either to humans or to the environment), the proposal cannot be considered as an
extension to a low-risk project. Any requirement for higher levels of containment precludes
the proposal being considered as an extension - eg, if the project is elevated to Class 2 or
Class 3, or if it includes the use of a GMHO that poses greater risks to human health and
Revised July 05
SG15(21)
Contained Use of Genetically Modified Organisms
_________________________________________________________________________________
_
safety ("non-ASIC") than its non-modified parent. Such proposals are regarded as new, and
will be subject to assessment by a technical sub-group of SCBS. If approved, the proposals
must be notified to HSE. (Section 9.6, below).
9.4
New or extended projects involving higher risk activities
Applications for higher risk projects must be made to SCBS (via the Health & Safety Services
office) on Form GM1Std. The proposals will be considered by a technical assessment group
of SCBS, which would be convened to meet at a mutually convenient time. This would
normally be not less than 2 weeks before a scheduled meeting of SCBS, to allow SCBS to
consider the report and recommendations of the group. Submissions made after this date
would be deferred to the next meeting of SCBS, with the exception of applications made
between the beginning of June and the end of August, when different arrangements apply
(Section 9.1).
Proposers should consider the impact of possible delays due to the notification process on the
planned starting date for the work (Section 9.6). Depending upon the class of activity and the
date of submission, there could be a delay of up to 3 months between submission and final
approval to start work. Proposers are therefore advised to plan well in advance to ensure that
their proposals are considered by the next available meeting of SCBS. Dates of meetings are
published in the University Calendar (Section 5.4). Items for the agenda of SCBS, together
with supporting papers, are normally circulated to members at least a week before the
scheduled date of the meeting.
9.5
Technical assessment groups
A technical assessment group will be assembled on an ad hoc basis to consider each higher
risk project as it arises, and would normally consist of:
•
the BSO or one of his deputies;
•
the SCBS Secretary;
•
the project proposer;
•
a representative of the staff to be involved on the project;
•
the HASC (or other representative of the local safety management); plus
•
appropriate independent technical experts unconnected with the project.
The technical experts may be either regular SCBS members, or be co-opted to the group
where SCBS members themselves did not have sufficient expertise in the relevant technical
areas. In some circumstances, one or more lay members of SCBS may be invited to join the
group, especially where the implications may be wider than the purely technical aspects that
are to be considered.
The technical group may recommend that the project be submitted to SCBS members for
formal approval, or that the application should be revised and resubmitted:
•
Following a decision to recommend approval, the proposals would normally be
submitted to the next meeting of SCBS for formal approval. Exceptionally, the postal
circulation route may be adopted (Section 9.1).
•
Proposers may be required to give a presentation to the meeting, in order to explain the
proposals in lay terms for the benefit of the non-technical members.
•
If the technical assessment group (or SCBS) rejects the proposals in their present form,
the revised proposals must be resubmitted and reassessed in the normal way.
SG15(22)
05
Revised July
Contained Use of Genetically Modified Organisms
9.6
Notification of Higher Risk Projects
All projects involving higher risk activities (Class 2 or Class 3 for GMMs, or those involving
non-ASIC GMHOs) must be notified to HSE in advance of starting work.
In addition to notification of new projects, "significant changes" to existing projects must be
notified. The term "significant change" may be interpreted as where there is a significant
change to the work which may have an effect on the risks of the activity - for example, where
the existing risk assessment does not cover the proposed additional work. Even if the class of
activity is not affected by the proposed change, the need for a new or significantly revised
risk assessment acts as the trigger for notification. For example, the insertion of a different
gene into a host organism would probably trigger a new notification, as would the use of a
different host. However, experience has shown that HSE may be prepared to discuss whether
such changes are notifiable on a case-by-case basis, especially where the changes lead to a
lowered risk. Project Supervisors should discuss their proposals with the BSO at the earliest
opportunity if they think their proposals fall into this category.
Formerly, "significant changes" could be notified without charge, but with the advent of the
new regulations, a full notification fee will be charged (see below). The only exception to this
is where the changes are purely of an administrative nature (for example, changes to the
composition of the GMSC, or notification of the closure of a project), when no fee is charged.
HSE acknowledge the receipt of each notification (Section 9.7).
Each notification must include "specified details" and supply "specified information" (ie, as
specified in the appropriate Schedules to the Regulations). A copy of the risk assessment
(plus any comments made by SCBS on the risk assessment) must accompany the notification.
The information given must include an indication of the degree of kill achieved by the
procedures used to inactivate waste, and a description of the methods used to monitor this.
(This is to comply with the requirement to "inactivate waste by validated means" (Table 2,
Schedule 8 to the Regulations)).
The University Standard Project Application Form (ref GM1Std) is designed to direct
applicants to supply the information required by HSE. Information may be transferred to the
appropriate sections of the HSE notification form CU2, or (if appropriate) a copy of the
University project application form sent to HSE. SCBS will approve the final wording on the
notification before it is submitted to HSE. A copy of the risk assessment must be sent to HSE
in conjunction with the notification.
A fee is charged for each new notification made. At the time of this revision (July 2005), the
fees are:
•
£829 for Class 2;
•
£899 for Class 3 (first notification; £864 thereafter)
•
£829 for harmful GMHOs; and
•
£622 for notification of "significant changes" under Regulation 15(3)), with the
exception of “administrative” changes, for which no charge is made.
The fees are reviewed on a regular basis, so applicants should check the current charge via the
BSO at the Health & Safety Services office, or via the H.S.E. website on
http://www.hse.gov.uk/biosafety/gmo/acgm/acgmwarn.htm
Once HSE have responded (whether this be an acknowledgement, or the issue of a Consent)
at the Health & Safety Services office, copies of the response would be sent to the Project
Proposer (subsequently referred to as the Project Supervisor), the Head of School and the
Revised July 05
SG15(23)
Contained Use of Genetically Modified Organisms
_________________________________________________________________________________
_
Health & Safety Co-ordinator. If HSE decline to give their Consent, the reasons for refusal
would be communicated only to the Project Proposer.
9.7
Commencing work on projects
Prior to work starting on an approved project, the Supervisor should:
•
note, and comply with any particular conditions placed on the approval, either by the
BSO, SCBS, or by HSE (see below);
•
require that all new workers who are recruited to join the project obtain a copy of this
guide; and then
•
ensure that all relevant staff have received written approval from the UOHMA that they
may commence work on the project.
When work may physically commence depends upon the type of project:
• Work on "low-risk projects" may commence under BSO approval as noted earlier.
• For work on "higher-risk projects", the project Supervisor must await confirmation from
the Health & Safety Services office that the work may start. Confirmation will only be
given following compliance with all relevant legal requirements.
• Class 2 activities
Work may commence immediately following receipt of the official acknowledgement of the
notification by HSE. However, if the Proposer wishes to claim that some of the information
contained in the application is confidential (and should not be disclosed on the Public
Register), he/she is advised to wait until HSE has decided whether to accept the claims before
starting work (Section 10).
• Class 3 activities
Work must not commence until HSE has given its consent. For a "first use" of Class 3
activities, HSE must inform notifiers whether or not consent has been issued within 90 days
of acknowledging receipt of the notification. For subsequent Class 3 activities, the period is
45 days. If, during the course of the assessment procedure, HSE decide that they need
additional information to evaluate the proposal, they will "stop the clock" ie, the time between
making the request and the supply of the requested information is not counted as part of the
specified period.
Note: Although Class 3 activities were undertaken under the previous Contained Use
Regulations, there were none in progress when the new Regulations came into force. The
University is therefore required to submit a "First Use" notification for the first new Class 3
project submitted after 15 February 2001. This requirement has the effect of imposing the "up
to 90 days" period between notification and the issue of a Consent. Subsequent Class 3
notifications will revert to the 45 day period.
• Work with harmful ["non-ASIC"] GMHOs
Work may only commence 45 days after the information has been notified to HSE, whether it
be a first or subsequent use. HSE Consent is not required.
9.8
Review of projects
It is a requirement of the Regulations that there must be a regular review of all GM risk
assessments, with the local GM Safety Committee (i.e., SCBS in the University) having the
duty to advise on those risk assessments. Although the frequency of review is not specified in
SG15(24)
05
Revised July
Contained Use of Genetically Modified Organisms
the Regulations, following a recent inspection of GM facilities at the University, HSE
recommended that an annual review be undertaken. One particular concern was that projects
could go outside the bounds of an existing risk assessment, and not be properly reviewed,
either by the project supervisor or by the local GM Safety Committee.
In order to effect this recommendation:
•
All active GM projects must now be reviewed on an annual basis;
•
Where appropriate, an updated Lay Summary must be submitted to SCBS for comment
and approval at their meeting in January of each year. (If there have been no changes
since the previous Lay Summary was submitted, a statement to that effect must be made
to SCBS for the January meeting.)
•
Failure to submit either an updated Lay Summary or a confirmatory statement (above)
may result in the project being declared "inactive" by SCBS, with storage of GMOs
being the only permitted activity.
•
Where a project is "inactive", a project supervisor will be required to sign a declaration
to that effect and submit it to the January meeting of SCBS.
•
If a project supervisor wishes to reactivate an inactive project, a revised risk assessment
and Lay Summary must be submitted for approval by SCBS at one of its scheduled
meetings.
9.9
Information, instruction and training
All GM workers should be aware of the requirements of the law relating to Contained Use of
GMOs. All new GM workers are therefore expected to attend either of the two training
seminars on the law relating to contained use of GMOs - see the "Training" page of the
Health & Safety Services office Web Site
(http://www.fmd.rdg.ac.uk/safety/training.asp)
for details.
New GM workers should also be referred to all relevant HSE and University guidance. In this
context, the term "worker" includes University teaching and technical staff; post-doctoral
workers, research assistants and postgraduate students, whether they are employed by the
University or any other body, or are grant-funded. (See Safety Note 16 [via
http://www.fmd.rdg.ac.uk/safety/documents/SafetyNotes/NOT16StudsuperGM.doc ] for
procedures applicable to students on taught courses involving GM work).
If a worker is new both to the School, and/or to GM work, he or she may need to be trained in
the techniques to be used. All training given should be fully documented, for example,
training in practical techniques should be recorded in the Supervisor's project file as well as in
the worker's laboratory notebook. For projects involving higher hazard organisms, the
specific details of the training should be fully documented as a formal training record.
All workers on the project should also be familiar with and comply with their Local Rules
(Section 7 of Safety Guide 14).
10
CONFIDENTIALITY ISSUES
10.1 Overview
Intending applicants should be aware that all the information (with the exception of personal
information) contained in a notification to HSE is potentially disclosable to the public, in the
form of an entry in the Public Register relating to Contained Use work. HSE intend to allow
electronic access to this Register via the Web.
Revised July 05
SG15(25)
Contained Use of Genetically Modified Organisms
_________________________________________________________________________________
_
The areas for which disclosure may have the most serious implications are those of
intellectual property rights (patent applications, etc)., or where the proposal is being
conducted in conjunction with a company that claims commercial-in-confidence status for
some of the materials or information used. Other grounds for withholding information from
the Public Register include the possibility of compromising national security, or public order.
With the exception of proposals involving animal work (where security issues are an
important consideration), these other issues are not expected to feature strongly in
notifications from the University, and are not dealt with here. For more details, contact the
BSO.
Where confidential status is to be claimed on the grounds of potential damage to intellectual
property rights, the claimant is strongly advised to seek professional advice (eg, from a patent
agent) before submitting the project application to the Health & Safety Services office.
10.2 Claims for Confidential Status
If an applicant wishes to claim confidential status for any of the information contained in the
University project application form, he/she is asked to tick the appropriate box on the form,
and indicate the areas of the form for which that claim is made. The applicant is then asked to
sign the appropriate statement in Section 3 of the form. Applicants should be aware that their
intellectual property rights are safeguarded by SCBS: all SCBS members and independent
assessors have signed an agreement restricting their discussion and use of any information
contained in sections of the form claimed as confidential.
Confidentiality claims will only be relevant to higher risk activities, as low risk activities are
not notified to HSE. Low-risk proposals stay within the University during the assessment and
approval process, and are not therefore regarded as being in the public domain. By contrast,
where the activities are higher risk, they must be notified to HSE. They will enter information
(as specified in Schedules 5 and 6 to the Regulations - see Reference 1) onto the Public
Register within 14 days of receipt, unless a confidentiality claim is made. If such a claim is
made, those details for which the claim is made are withheld from the Register until HSE has
decided whether or not to grant the claim. Each claim for confidential status must be fully
justified. Note that some information cannot be claimed as confidential, for example, the
location of the intended activity. This is given as "The University of Reading" for all
University notifications, as The University is notified as a single set of premises.
10.3 HSE Response
If HSE decide that the claims are not to be granted, the project details will be entered onto the
Register 14 days after that decision is communicated to the applicant. This delay gives the
applicant time to withdraw the application if he/she so wishes. If the application is
withdrawn, the confidential information will not be entered onto the Register, and any
information already entered will be removed. The application may only be withdrawn if
work has not already commenced on the project.
Because work on a Class 2 project may commence immediately after HSE has acknowledged
its receipt (Section 9.7), if the proposer has commenced work on the project before receiving
a decision on the confidentiality claim, the details will be entered onto the Public Register.
Proposers are therefore advised to wait for a decision on the claims before starting work.
The situation is probably more straightforward for Class 3 proposals. HSE must give its
consent before work may begin, and any claims for confidential status would probably be
considered during the 45 days allowed before a Consent is issued.
HSE have advised that they wish to see claimants indicate the extent of information that could
be disclosed without damage to their interests. If HSE accepts a Confidentiality claim, they
SG15(26)
05
Revised July
Contained Use of Genetically Modified Organisms
will place an edited version of the information onto the Public Register. Prospective
Supervisors should therefore also prepare an edited version of their proposals for submission
to HSE (via the Health & Safety Services office).
HSE will regularly review each claim to determine whether it is still justified. For example,
the grant of a patent would result in withdrawal of any confidentiality claims that were based
on preservation of intellectual property rights. An appropriate entry would then be made on
the Public Register.
11
SUPERVISION OF STUDENTS UNDERTAKING GM WORK
Full details are given in Safety Note 16, and only a summary is given here.
There are two situations where undergraduates may be involved with GM work: when they
are participating in fully supervised laboratory practical classes, or an undergraduate research
project that involves the use of GMOs. These situations are treated differently in respect of
the registration of the individuals as GM workers. In both situations, anyone taking part in a
supervisory capacity - for example, as a demonstrator in a fully supervised practical class; or
as a GM research Project Supervisor - must be properly registered as a GM worker.
All supervised practicals that include GM activities must have been approved by the SCBS as
suitable for such use: they must all be in the lowest risk Class, ie, Class 1 (or equivalent).
Once the project has been approved, a list showing the names of the
Supervisors/demonstrators should be sent to the Health & Safety Services office before the
series of practicals starts. These details should include information on the experience of the
Supervisors and demonstrators in GM techniques. As the details of student numbers, etc., will
change from year to year, the project proposer should state the minimum levels of
demonstrator or Supervisor numbers that will be required, and their required level of
experience in GM techniques.
Undergraduates may also participate in approved GM projects in their School, or could (with
appropriate guidance from a prospective Supervisor) suggest their own project.
Normally, undergraduates may only be involved with Class 1 GM projects (or equivalent for
higher organisms). If a Project Supervisor wishes to allow undergraduates to work on Class 2
projects, he/she must submit a case to SCBS for approval. If SCBS were to approve, any
conditions they apply must be adhered to by the Supervisor. Any students permitted to work
on such an approved project must be demonstrably competent in basic "good microbiological
practice" techniques, and be subject to a high level of supervision. They must be subject to
the full health registration and surveillance procedures.
12
REFERENCES
The following publications (or later editions or revisions) are available for reference or loan
from the Health & Safety Services office (extension 8888).
1.
A Guide to the Genetically Modified Organisms (Contained Use) Regulations 2000.
ISBN 0-7176-1758-0, HSE Books
2.
HSE/ ACGM Compendium of guidance. ISBN 0 7176 1763 7 (Copies of relevant parts
of
this
guidance
are
issued
to
each
project
Supervisor).
Also available as a free download from HSE Health Directorate web site
[http://www.hse.gov.uk/biosafety/gmo/index.htm], or from HSE Books
Revised July 05
SG15(27)
Contained Use of Genetically Modified Organisms
_________________________________________________________________________________
_
13
3.
The Control of Substances Hazardous to Health Regulations, 2002 (as amended).
Approved Code of Practice; L5, Fifth edition, 2005, ISBN 0 –7176-2981-3, HSE Books
4.
The Management of Health and Safety at Work Regulations, 1992 (as amended).
Approved Code of Practice, C500 5/93. ISBN 0 –11-886330-4, HMSO.
ACRONYMS
The following acronyms and abbreviations have been used in this Safety Guide.
ACOP
ACDP
ACGM
HASC
ASIC
ASTA
BSO
COSHH
DEFRA
DNA
GM
GMHO
GMM
GMO
GMSC
GP
HSC
HSE
MAFF
MOHSAW
Non-ASIC
RNA
SCBS
UOHMA
SG15(28)
05
Approved Code of Practice
Advisory Committee on Dangerous Pathogens
Advisory Committee on Genetic Modification
Health & Safety Co-ordinator
"As Safe in Containment" (as the non-modified parent)
Health & Safety Technical Advisor
Biological Safety Officer
Control of Substances Hazardous to Health (Regulations)
Department of the Environment, Food and Rural Affairs
Deoxyribonucleic acid
Genetic Modification also Genetically Modified
Genetically Modified Higher Organism
Genetically Modified Microorganism
Genetically Modified Organism
Genetic Modification Safety Committee
General Practitioner
Health and Safety Committee
Health and Safety Executive
Ministry of Agriculture, Fisheries and Foods (now DEFRA)
Management of Health and Safety at Work (Regulations)
"Not as safe in Containment" (as the non-modified parent)
Ribonucleic acid
Sub-committee for Biological Safety
University Occupational Health Medical Advisor
Revised July
The University of Reading Safety Information Sheet
SPECIMEN PROJECT APPLICATION
FOR LOW RISK GM PROJECT
SG15 Appendix 1
This specimen application and risk assessment presents a hypothetical application to
undertake work with inherently safe genetically modified microorganisms (GMMs), and also
with "ASIC" genetically modified higher organisms (GMHOs) i.e., those that meet the criteria
for "posing no greater risks to humans than their non-modified parents". The application is
made on the "low risk" project application form GM1LR2002 coupled with the risk
assessment form GM03.
The application would initially be assessed by the BSO to check that it conforms with the
criteria for low risk projects. If he agrees with this assessment, it would be sent to an
independent technical assessor, who would be asked to comment and make a recommendation
whether to accept or reject the proposal.
Assuming that the assessor recommends acceptance, the BSO would give temporary approval
in writing to the project supervisor for the project to commence. The temporary BSO approval
would be subject to ratification at the next available meeting of SCBS: a copy of the
"statement of overall risks" and the "Lay summary" would be presented to the sub-committee,
and form the basis of discussion. Any decision of the sub-committee would be communicated
in writing to the project supervisor: if the temporary approval were not ratified, work would
have to stop and the project resubmitted to the next meeting of SCBS. Any resubmitted
project would not then be eligible for BSO approval.
All workers named on the project application would have to register for this project - new
workers (not previously registered for GM work at the University) would have to submit the
completed GM worker registration form (GM02) to the BSO. They would also have to submit
a copy of the registration form to the Health & Safety Co-ordinator and the project supervisor.
The BSO would issue an "approval to commence work" letter, and work could commence on
the project. The duration of the worker approval would be limited to the period stated in the
original application form, unless a request is made to extend the approval period. A condition
of registration is that each worker attends a training session on the requirements and
procedures relating to GM work.
___________________________________________________________________________
Revised July 05
SG15 Appendix 1(1)
Specimen project application for low risk GM project
_________________________________________________________________________________
_
Section 1: Project information
1.
2.
Area or Department where the work
is to be carried out.
Please delete as appropriate
Department of Science Studies
Project Supervisor, i.e.,
Member of staff responsible for
directing the work.
Clone
Title Dr.
Surname
I.M.A.
Initial
5678
Extension
[email protected]
e-mail
3.
Staff involved operationally on the
project,
and details of their qualifications/
experience in GM techniques. If
appropriate, give numbers of
previous
GM
projects.
Name
Position
Epsom, J.
Postdoctoral
fellow
Postgraduate
Lecturer
Technician
Fraser, G
Clone, I.M.A.
Doitall, I.
For undergraduate student
classes or projects, please tick
box, and attach separate lists of
participating
students
and Undergraduate class?
demonstrators; appropriate lists
should be resubmitted each year
the project is run.
Qualifications/
GM experience
B.Sc.; Ph.D. 15 yrs.
GM
B.A.
No
GM
D. Phil. 15 yrs.
HNC; 3 yrs. GM
See also Note 8 on p2.
4.
Number / location, and containment
level of laboratory facilities
proposed for project.
Please tick box if any of these
facilities are to be used for GM work
for the first time.
Room number/ location
102
103
Containment level
2
2
First use?
5.
Please tick box and give reference
number
Extension?
Previous Ref. No.
if this project is to be considered
as an extension to a previouslyapproved project.
Please then complete appropriate sections in the
remainder of this form, so as to emphasise the connection
between the original project and this application.
Note that this form must NOT be used to request an
extension to an existing Class 2 or Class 3 project.
6.
Title of project
Modification of potato tubers with the aim of
improving crisping qualities.
___________________________________________________________________________
SG15 Appendix 1(2)
Revised July 05
Specimen project application for higher risk (Class 3) GM project
7.
Purpose of the project,
stated in layman's terms. This The aim of the project is to modify the carbohydrate
statement may be incorporated metabolising enzymes in potato tubers with a view to
preventing the formation of glucose and other reducing
into the Project Lay Summary.
sugars during low-temperature storage of potato
tubers. Potatoes are normally kept in cold stores after
harvest prior to processing (which could be for several
months.) The presence of reducing sugars in the slices
of potato during frying for crisp manufacture results in
blackening of the crisps - leading to rejection by
quality control, and significant losses during the
manufacturing process. The project aims to firstly
identify, then inactivate the enzymes responsible.
Gene inactivation will be by antisense gene
technology: gene sequences designed to be
complementary to the genes will be inserted into
potatoes, and screened for their effects on reducing
sugar accumulation during tuber storage.)
___________________________________________________________________________
Revised July 05
SG15 Appendix 1(3)
Specimen project application for low risk GM project
_________________________________________________________________________________
_
8.
The scope and expected results of
the project.
(Where relevant: if the project
consists
of
a
connected
programme, please describe the
expected
scope
of
the
programme.)
The project will examine the possibility of inhibiting
the production of reducing sugars in potato tubers
during cold storage. The enzymes which may be
responsible for this event include one or more of:
α-amylase;
invertase;
ATP-dependent
phosphofructokinase and pyrophosphate-dependent
phosphofructokinase.
Total messenger RNA pools from stored potato tubers
(cultivars Record and Saturna, commonly used for
crisp manufacture) will be isolated, and cDNAs
prepared and amplified by reverse transcriptase PCR.
The cDNAs will be used to create a library in
LambdaGEM (or similar) vectors, which will be
screened for enzyme activity in appropriate disabled
strains of E. coli (complying with “inherently safe
criteria”) by Western blotting against monoclonal
antibodies to the purified potato enzymes.
Clones carrying the appropriate enzymes will be
multiplied up and the inserted DNA sequenced to
obtain information for synthesis of the respective
antisense sequence of each gene. (When transcribed,
the antisense sequence will produce an RNA sequence
that will bind to and inhibit the translation of the
enzyme messenger RNA.) Each synthetic antisense
sequence will be inserted into the binary plasmid
pBIN19 and replicated in E. coli, before being
transferred to the disabled Agrobacterium strain L4404
for insertion into potato cells under the control of a
cold-activated tuber-spcific promoter. The marker
genes GUS and NPTII (Kanamycin resistance) will be
used to help control the process and select the desired
transformed cells. Transformed potato cells will be
grown in suitable media to produce potato plants
transgenic for the desired antisense sequences.
Transgenic potato plants will be grown in containment
glasshouse, and tubers will be screened by testing
accumulation of reducing sugars during lowtemperature storage. Plants showing greatly reduced
accumulation of reducing sugars will be further
screened for commercial development potential.
There are no plans at present to initiate a “deliberate
release” by applying for permission to grow these
modified potatoes in the open field: all growth of
modified plants will be undertaken in a growth room
or in a containment glasshouse.
___________________________________________________________________________
SG15 Appendix 1(4)
Revised July 05
Specimen project application for higher risk (Class 3) GM project
Section 2: Details of Genetic Modification
Please complete
Part A for GMMs, and / or
Part B for GMOs that are not microorganisms.
You will need to complete both parts of this section if your project involves
both GMMs and higher organisms.
[Remember: cell cultures are classified as microorganisms.]
Please delete part A if not applicable
Part A: Activities with genetically modified microorganisms based on
“inherently safe” donors, vectors and recipients.
Data or references to support the "inherently safe" nature of the above may be required.
This section provides the basic information for confirmation of the activities as “low risk”.
y
y
Please give sufficient detail to allow the independent assessor to confirm your risk
assessment. Include all parental / donor and recipient organisms (or indicate the
possible range, identifying the elements of greatest potential risk), and each host-vector
system to be used. Use one entry for each separate system in each heading. Include
supporting references where appropriate.
If GMMs are to be imported into the University from elsewhere, the information on
“construction of the GMM” must be obtained from the prospective supplier. [N.B.: it is
permissible to use a risk assessment prepared by the supplier, provided that this gives
the information required in the headings below.] Such GMMs must NOT be
obtained before obtaining project approval.
Please tick box if the GMO was constructed elsewhere
and is being brought onto University premises
9.
The identity, source and function
of each sequence of genetic
material to be inserted.
Identify all potentially harmful characters
such as toxin genes, or other pathogenic
functions.
[Please extend each section as necessary to
include all donor or parental organisms, etc. use one heading for each insert.] State ACDP
or other hazard categorisation [e.g., DEFRA]
of any donor or parental microorganism.
Where relevant, identify any antibiotic
resistance markers and potential for
transmission; mutations attenuating virulence
and possibility of reversion; identify all
sequences which may affect expression of
genes in the recipient microorganism.
See Note 4 on p. 2
Â
(a) cDNAs initially corresponding to total mRNA
pool of stored potato tubers (Solanum
tuberosum, cvs Record and Saturna.), produced
by RT-RCR amplification of mRNA pool.
(b) Antisense constructs corresponding to
enzymes α-amylase; invertase; ATP-dependent
phosphofructokinase and pyrophosphatedependent phosphofructokinase from potato
tubers as above. (Constructs to be prepared
following cloning/ sequencing of cDNAs of
these enzymes.)
(c) β-glucuronidase [GUS] and Neomycin
phosphotransferase II (NptII) marker genes for
control purposes
___________________________________________________________________________
Revised July 05
SG15 Appendix 1(5)
Specimen project application for low risk GM project
_________________________________________________________________________________
_
10. Identity of all recipient
microorganisms.
If you wish to use a range of
possible recipients, please identify
the extent of the range, and give
justification for them to be
considered as part of the same
project. See ACGM Compendium
of Guidance for information on
disabled bacterial host strains.
For cell cultures, give strain/ line
information as well as species.
Use one line for all strains of each
species of recipient (with the
same level of disability. See Note
6 on p. 2
11. Identity of the vector(s)/ vector
family(ies).
State whether vectors are mobilisable;
identify any antibiotic markers present.
See Note 5 on p. 2. Where vectors are
not used, state means of insertion of
DNA into the recipient (e.g., use of the
“gene gun”.)
(a) Disabled strains of E. coli, complying with
inherently safe criteria. (such as DH5α, and
XL-1)
(b) Disabled Agrobacterium tumefaciens, strain
L4404 (or equivalent
a) LambdaGEM-11 cloning vector (or equivalent)
for initial cloning of cDNAs into E. coli, and
derivatives used for direct sequencing of
inserted DNA in appropriate identified clones.
b) pBIN19 (or functionally identical disarmed Ti
plasmids) carrying inserted synthetic DNA
sequences (corresponding to the antisense
sequences of the genes of interest) under the
control of the tuber-specific cold-activated
promoter prST1, capable of replication in both
E. coli and Agrobacterium (GUS and NptII
genes under the control of the pNos (nopaline
synthase) promoter.)
___________________________________________________________________________
SG15 Appendix 1(6)
Revised July 05
12. Nature of expected properties of
the GMM.
(Take into account “insertional”
effects; expression of the insert
and/or genes comprising the
vector, vector replication; stability
and/or maintenance of the vector
in the recipient; potential for
transmission of genes to other
organisms, survival in the
environment, etc.) Do not forget
“marker” genes such as antibiotic
resistance
genes
and
the
possibility
of
compromising
antibiotic treatment.
.E. coli strains not expected to be harmful either to
humans or the environment, as based on disabled
laboratory strains; inserted genes not expressed in E.
coli, and cannot survive outside of specialist media.
Agrobacterium strains carrying disarmed plasmids
with inserted genetic sequences could transfer genes
to recipient plants if the organisms escaped from the
laboratory. Antibiotic resistance and other marker
genes widespread in the environment, and no impact
expected even if the Agrobacterium were to escape
and become established in the environment. Transfer
of cold-activated carbohydrate metabolism genes to
plants other than potato unlikely to occur, as genes
under contol of potato-tuber specific promoter. If the
genes were transferred to potatoes in the
environment, it could reduce ability of tubers to
survive cold conditions.
13. State maximum culture volume to
be used at any one time.
500 ml of E. coli; 250 ml of Agrobacterium
14. Identify all types of operation with
potential for dispersal. .
Centrifugation/ resuspension of centrifuged cultures
(For example, aspiration;
resuspension of cultures;
centrifugation or sonication).
15. Potential for exposure, based on
maximum culture volume to be used;
type of operation and potential for
dispersal
16. State methods to be used for
decontamination of waste and
equipment. If disinfection to be
used, identify disinfectant and state
conditions of use. Identify any
methods used to monitor
effectiveness of disinfection.
Low/ Negligible
All waste to be autoclaved (121o, 20 min) before
discard; equipment disinfected using Hycolin
phenolic disinfectant (1%) for minimum of 2 hours;
recycylable glassware soaked in Chloros
disinfectant [2500 ppm available Chlorine] for
minimum 2 hours before collection for washing and
re-sterilisation. No surviving bacteria detected
under conditions of use; effectiveness of
disinfectant monitored by checking available
chlorine levels under “in use” conditions on weekly
basis.
___________________________________________________________________________
Revised July 05
SG15 Appendix 1(7)
Specimen project application for low risk GM project
_________________________________________________________________________________
_
Go to Risk Assessment Form, GM03 to assess the risks, based on:
(1) Any possible hazards associated with
•
•
•
the donor or sequence of DNA to be inserted;
the recipient organisms, and
the vectors to be used.
(2) The potential for exposure estimated above.
17. Based on your answers in Q1 Q5 of the risk assessment form,
does the GMM have any capacity
for to cause harm to humans or
to the environment?
No (please delete as applicable)
If “YES”, you should complete the full project
application form, GM1Std, available via the “Forms”
page of the Health & Safety Services web-site,
http://www.fmd.rdg.ac.uk/safety/forms.asp?DSID=12
Part B: Projects involving the use of genetically modified higher
organisms. (Please delete this Part if not applicable)
References or data may be required to support the non-harmful status of recipient or donor
organisms.
Please tick box if the GMO was constructed elsewhere
and is being brought onto University premises
18. Identity of all recipient organism(s).
Give common and scientific names, and,
where relevant, strain, cultivar or subspecies
designations.
19. Where applicable, the identity of the
host/vector system, or the method
used for the genetic modification.
Potato (Solanum tuberosum), cvs. Desirée;
Record and Saturna (cells modified as in Part
A, and intact plants grown from tissue culture.)
Transformed plantlets grown in presence of
antibiotic, then screened to ensure absence of
Agrobacterium.
See Part A.
___________________________________________________________________________
SG15 Appendix 1(8)
Revised July 05
Specimen project application for higher risk (Class 3) GM project
20. Origin and intended function of
inserted genetic material.
See Part A.
If you wish to use a range of possible
gene donors, please identify the
extent of the range, and give
justification for them to be
considered as part of the same
project.
See Note 7 on p. 2
Identify any sequences that may
affect expression of genes in the
recipient organism.
Go to Risk Assessment Form, GM03 to assess the risks, based on:
(1) Any possible hazards associated with
•
•
•
the donor or sequence of DNA to be inserted;
the recipient organisms, and
the vectors to be used.
(2) The potential for exposure estimated above.
21. Based on your answers in Q8 - Q11 of
the risk assessment form GM03,
could insertion of the above genetic
material create any potentially
harmful effects in any of the
recipients?
No (please delete as applicable)
If “YES”, you should complete the full project
application form, GM1Std, available via the “G”
page of the Health & Safety Services web-site,
http://www.fmd.rdg.ac.uk/safety/forms.asp?DSID=
12.
___________________________________________________________________________
Revised July 05
SG15 Appendix 1(9)
Specimen project application for low risk GM project
_________________________________________________________________________________
_
Section 3: Declarations
[If the form is completed by computer and returned by e-mail, this and the following page
should be printed off, signed, and returned by internal mail to the Health & Safety Services
office marked “For the attention of SCBS”.]
1. Proposers' Declaration:
I am submitting this application for approval at the next meeting of the Safety Subcommittee for
Genetic Modification. I agree that work will not commence on this project until SCBS has given its
approval, and that all workers will be properly registered with the Occupational Health Medical
Adviser. I also enclose a copy of the risk assessment form GM03.
Name (please print or type)...........I. M .A .Clone.........................................
Signed........................................................................Date ........29 /02/02..............
2. Supporting Declaration
(The person supporting this proposal must not be involved in the project being proposed.)
I support the presentation of this proposal to the subcommittee.
Name (please print or type)
A. Brown...................…….........
Signed ..................................................................Date ..... ........29 /02/02...........
Status in organisation:
Health & Safety Co-ordinator
(please delete as necessary)
•
•
•
PLEASE CHECK THAT:
ALL RELEVANT SECTIONS OF THIS FORM HAVE BEEN COMPLETED,
A COMPLETED RISK ASSESSMENT FORM (GM03) IS SUPPLIED, AND
ALL DECLARATIONS ARE SIGNED.
Proposers must deposit a copy of both completed, signed forms with the Health & Safety
Co-ordinator/ Health & Safety Technical Administrator, as well as keeping copies for
their own records.
Please Note:
Proposals giving insufficient information may be subject to delay if the forms are
returned for the proposer to supply more information.
BSO approval is subject to ratification by a full meeting of SCBS, who may attach
conditions to their approval.
Failure to ratify the BSO approval will result in the project being declared inactive.
Work may then only recommence once SCBS has given its approval to a revised project
application.
___________________________________________________________________________
SG15 Appendix 1(10)
Revised July 05
Specimen project application for higher risk (Class 3) GM project
Reminder:
All workers named in this proposal must also complete and return a worker registration form
[ref. GM02] to the Occupational Health Medical Adviser.
The worker registration form(s) should only be submitted after the Project Supervisor has been
notified in writing that the project has been approved.
Worker Approval is subject to any conditions imposed by SCBS when the project is approved.
An individual may not start work on the project until written confirmation has been received
stating that his/her registration has been approved.
For committee use only.
Date of
submission........................................................................................................
Independent assessor……………………. ....................................................
Comments received by (date) ............................................
BSO Approval given?
Y/N
(date)...........................................
(Signed).........................................................................................................................
.
Confirmed by full committee meeting on
(date)....................................................................
Decision: Approved /Rejected/ More information required/additional conditions
imposed. (delete as necessary)
(signed) ....................................................................Date............................
Nature of any special conditions
imposed?:......................................................................................................................
......................................................................................................................................
......................................................................................................................................
......................................................................................................……………………....
................................…………………………………………………………………………..
___________________________________________________________________________
Revised July 05
SG15 Appendix 1(11)
Specimen project application for low risk GM project
_________________________________________________________________________________
_
Risk assessment
Where this form is used for the Annual Review of the risk assessment,
highlight any changes made since the last risk assessment form for this
project was submitted.
Please complete
Part A for GMMs, and / or
Part B for GMOs that are not microorganisms.
You will need to complete both parts of this section if your project involves
both GMMs and higher organisms.
Please delete part A if not applicable
Part A: Activities with genetically modified microorganisms
Use the information in the project proposal form GM1 to answer the questions posed
below. You must justify any statements made, e.g., by references to the scientific
literature where necessary.
1.
Identify all
potentially
hazardous
properties of each
GMM
Include hazards to
human health and
safety, and hazards to
the environment;
consider properties of
host, vector, insert, and
of final GMM. Use one
entry for each type of
GMM.
See Note 2
E. coli strains are disabled, and cannot survive in the
environment or colonise the human body, the inserted genetic
material is not expressed in E. coli. No hazards to human health
or to the environment.
Agrobacterium strains are non-pathogenic to humans, and are
disarmed, thus non-pathogenic to plants. They could survive in
the environment, and transfer the inserted genetic material to
plants. (This material is either the antisense version of genes
involved in carbohydrate metabolism, or one or more marker
genes.) If an escape from containment were to occur, resulting in
transfer of genetic material to susceptible plants, there is a
possibility that marker genes (including resistance to the
antibiotic kanamycin) or the carbohydrate metabolism anti-sense
genes could be expressed in plants. In this case, the only plants
that could be affected would be potatoes, as the genes are under
the control of a potato-specific (cold-activated) promoter.
___________________________________________________________________________
SG15 Appendix 1(12)
Revised July 05
Specimen project application for higher risk (Class 3) GM project
2.
Likelihood of
hazards associated
with GMM being
realised.
(Take into account any
disabling mutations in
the GMM; the scale and
type of operations, plus
the containment level
to be used.)
Likelihood of E. coli causing any harm = Negligible.
Likelihood of Agrobacterium escaping from containment and
affecting potatoes in the environment = Negligible.
Maximum culture volume to be used = 500 ml of E. coli; 250 ml of
Agrobacterium
Use the terms "High",
"Medium", "Low" or
"Negligible" and give
details of scale and
type of operations to
be done with each
GMM.
See Note 3
3.
Consequence of
hazards being
realised. (For the
purpose of this
assessment,
assume that the
GMM can colonise
or enter the
environment.)
Use the terms "Severe",
"Medium", "Low" or
"Negligible" and give
details of the expected
consequences
4.
Estimation of risk
magnitude
(Magnitude of Risk =
"likelihood"
x
"consequence" of causing
harm.)
Use one line for each
GMM.
For E. coli, consequence "negligible", as strains used are nonpathogenic, and cannot survive outside of specialist media.
For Agrobacterium, strains used are disarmed, and nonpathogenic towards plants. Consequence expected to be "low"
even if the Agrobacterium were to infect susceptible plants and
transfer their inserted genetic material, as this is likely to
decrease the ability of the transgenic plants thus created to
survive in the environment and compete with non-transgenic
l
Risk effectively zero for E. coli [negligible consequence;
negligible hazard].
Risk effectively zero for Agrobacterium [negligible likelihood
and low consequence]
See Note 5, "Risk matrix
5.
For each GMM
where the risk is
not "effectively
zero", identify the
nature of the "nonzero" risks.
N/A
See note 5
6.
Identify all
additional
measures
necessary to
control the above
risks.
(Containment level 1 sufficient to control all risks.)
___________________________________________________________________________
Revised July 05
SG15 Appendix 1(13)
Specimen project application for low risk GM project
_________________________________________________________________________________
_
7.
State class of
activity (equivalent
to that level of
containment with
characteristic
measures identified
as necessary in Q6)
See Note 1
Class 1
Note: Activities in Class 2 and/or Class 3 must be notified to HSE
[Please delete part B if not applicable]
Part B: Projects involving the use of GMOs that are not
microorganisms.
8.
Identify all
potentially
hazardous
properties of the
GMO(s)
Take into account any
toxic or allergenic effects
of expression in the
GMO; ability to transfer
genes to, or interfere with
other organisms in
established ecosystems;
acting as a novel
reservoir for a pathogen,
etc.
See Note 2 on p. 1.
Potatoes expressing antisense genes not expected to differ in
toxicity or allergenicity from wild-type potatoes, since
expression of antisense genes does not involve any novel
protein production, but blocking of expression of native mRNA
within stored potato tubers.
Potatoes expressing marker genes such as kanamycin resistance
or GUS will have proteins not normally found in potato tubers,
but the proteins involved are commonly found in the
environment, and would a normal component of the human
diet. No hazards to human health expected (and these could
only be expected if the potatoes were to be consumed.)
Consumer trials of crisps prepared from these potatoes would
not be undertaken without extensive preliminary toxicity
studies to confirm the lack of toxicity associated with the
tubers/ potato products.
Transgenic potatoes expected to have reduced ability to survive
in the environment due to blocking of normal carbohydrate
metabolism in the tubers.
Transgenic potato plants could flower and release GM pollen
into the environment, and fertilise receptive potato plants in the
environment.
9.
Estimate the
likelihood that
these hazards
could be realised.
Take into account the
containment measures to
be applied.
Use the terms "High",
"Medium", "Low" or
"Negligible" and give
details of scale and type
of operations to be done
with each GMO.
Likelihood = "Negligible", as controls applied to prevent
transgenic tubers from leaving the laboratory/ growth room/
glasshouse without being killed (all waste autoclaved.) All
plants will be regularly examined for incipient flower bud
formation; all flowers will be removed before they open to
prevent pollen maturation and dispersal.
___________________________________________________________________________
SG15 Appendix 1(14)
Revised July 05
Specimen project application for higher risk (Class 3) GM project
10. Estimate the
consequence of
these hazards
being realised.
(For the purpose of this
assessment, assume that
the GMO is not
contained.)
Consequence = "Low/ Negligible", in all cases. Using a "worstcase scenario"of transgenic potatoes entering the environment
around the laboratory/ growth room/ glasshouse, location is such
that no potatoes grown within 500m - thus such an escape would
not have any consequences for horticultural or agricultural
practices.
Use the terms "Severe",
"Medium", "Low" or
"Negligible" and give
details of the expected
consequences
11. Estimation of risk
magnitude
(Risk = "likelihood" x
"consequence" of
causing harm.)
Risk "effectively zero" [Negligible likelihood; low/ negligible
consequence.]
Use one line for each
GMO.
See Note 5, "Risk
matrix" on p. 1.
12. For each GMO
where the risk is
not "effectively
zero", identify the
nature of these
"non-zero" risks.
13. Identify all
measures
necessary to
control the above
risks.
14. Based on the
information in Q12,
does the GMO pose
any greater risks to
human health and
safety than the nonmodified parent?
See Note 4
N/A
N/A
No
(Note: the answer to this question is independent of any additional control
measures identified above.) If "Yes" the project must be notified to HSE.
___________________________________________________________________________
Revised July 05
SG15 Appendix 1(15)
Specimen project application for low risk GM project
_________________________________________________________________________________
_
Summary Statements
The main sources of information for "lay" members of the Committee are the
"Statement on overall risks" and the Lay Summary. They will be copied and
submitted to the Committee for formal approval. Sufficient detail, in non-technical
terms, should therefore be given to allow them to understand the aims of the project,
and to confirm the low–risk nature of the project.
Statement on the overall risks associated with this project
Use
the
information
provided in Parts A and/
or B to draft this
statement. You should
identify
the
most
hazardous elements of
the project, and any
procedures necessary to
control the associated
risks. Extend the section
as necessary.
No hazards to human health identified (and hence no risks), either
either from GMMs or from the transgenic potato plants. There is no
intention to undertake any taste trials on crisps made from the
transgenic potato tubers at this stage. Although this is a possibility
in the future, a separate application will be made to the Ethics
Committee for such work, and then only if toxicity trials confirm
the expected lack of toxicity of the tubers.
Any risk to the environment is expected to be negligible, as hazards
from the GMMs and the transgenic potato plants expected to have
negligible likelihood and low/ negligible consequence of being
realised. Containment level 1/ good glasshouse practice sufficient
to control all risks.
___________________________________________________________________________
SG15 Appendix 1(16)
Revised July 05
Specimen project application for higher risk (Class 3) GM project
Lay summary of the project
Outline the aims and
expected results of
the project. Where
possible, avoid the
use of acronyms and
abbreviations,
and
explain or spell out
any used. ("DNA" and
"RNA" need not be
spelt out.) Describe
the measures to be
put in place to control
the identified risks,
including those taken
to
decontaminate
waste
/equipment
and/or
prevent
dissemination
of
GMOs
from
the
laboratory..
[Expand
box
as
required.]
The aim of this “low risk” project is to improve the crisping qualities of
potato tubers kept in low-temperature storage: currently, tubers tend to
degrade some of the starch to reducing sugars during prolonged lowtemperature storage (“low temperature sweetening”). This causes a problem
when the tubers are processed for crisp production, as the presence of
reducing sugars causes blackening of the crisps, and rejection by quality
control. The project therefore aims to identify the enzymes responsible for
this effect, and to try to produce transgenic (genetically modified) potatoes in
which the synthesis of these enzymes is prevented.
Messenger RNAs will be extracted from potatoes, and used to create cDNA
copies (DNA copies of the messengers), which will be cloned into E. coli for
identification and subsequent sequencing. When this has been achieved, the
sequence information will be used to construct synthetic “antisense” copies of
the genes, which will be inserted into the potato cells with the aim of blocking
the function of the normal genes. The antisense genes are designed to produce
“antisense messengers” that will interfere with the function of the native
genes for these enzymes, and thus prevent the enzymes being made in the
tubers. They will be inserted in such a way that they will only be activated
and expressed in potato tubers during cold-storage.
All GMMs created will be based on organisms complying with “inherently
safe criteria”, and will only require containment level 1 (Class 1 activities
with GMMs.) As such, the GMMs will present minimal risks to human health
or to the environment. All waste will be autoclaved before disposal, and
glassware will be treated with hypochlorite disinfectant before washing and
re-sterilisation and re-use to minimise the possibility of accidental escape to
the environment.
The DNA constructs will be inserted into potato root-tip cells by the
Agrobacterium transformation technique. (Initially, the cultivar Desirée will
be used for the transformation, as it is easier to transform than “Record” or
“Saturna”, but once the conditions have been optimised, attempts will be
made to obtain direct transformation of the crisp-making cultivars.) The
transformed potato root cells will be grown in the presence of antibiotics to
destroy (and then shown to be free of) any Agrobacterium before any
subsequent treatment.
Transgenic clones of potato root-tip cultures will be identified and grown in
appropriate media to create whole potato plants transgenic for the antisense
genes. There are no plans at present to initiate a “deliberate release” by
applying for permission to grow these modified potatoes in the open field: all
growth of modified plants will be undertaken in a growth room or in a
containment glasshouse. There are negligible risks to human health or to the
environment posed by the transgenic potatoes. There are currently no plans to
undertake any taste trials with the tubers produced by the modified plants, and
all staff will be required to sign a declaration which recognises that these
tubers are not to be consumed.
___________________________________________________________________________
Revised July 05
SG15 Appendix 1(17)
Specimen project application for higher risk (Class 3) GM project
The University Of Reading Safety Information Sheet
SPECIMEN PROJECT APPLICATION
FOR HIGHER RISK (CLASS 3) GM PROJECT
SG15 Appendix 2
This specimen application and risk assessment presents a hypothetical application to
undertake work with a genetically modified Adenoviral vector, intended for use in the therapy
of acute myeloid leukaemia in adults. The application is based on the standard University
project application form, and is intended to demonstrate the level of detail that would be
required to satisfy SCBS. (Whether or not this would suffice for HSE is not known.)
An application of this type (which is classified as Class 3 as a "worst-case scenario") would
have to be considered by a technical group as part of the standard assessment process. The
group meet to hear a presentation made by the Project Supervisor (in this case, Dr. Clone),
who would be prepared to answer any questions on the project that members of the group
might have. Copies of papers cited in support of the application would be available for
perusal if required.
If the group were to recommend acceptance (with or without any modifications to procedures,
etc.), a report and recommendations would be submitted for consideration at the next
available meeting of SCBS. The report would be circulated with the Agenda papers, to allow
members to study the information before the meeting. Members could, if they wished (and
notified the Secretary of SCBS accordingly) request the prospective project Supervisor to
attend the meeting of SCBS to answer any queries raised on the report.
If SCBS were to approve, the project would be submitted to HSE, together with the
appropriate fee. (Information on the University project application form would be transferred
to the HSE form, CU2, plus any comments that SCBS might have on the risk assessment.)
Sections with confidential information would be clearly marked; an edited version suitable for
inclusion in the Public Register would also be prepared and submitted at the same time. The
claim for confidentiality would be on the basis of safeguarding intellectual property rights.
Work would not be able to commence of the Class 3 aspects until HSE had given its Consent,
although work on the Class 2 aspects could start once agreement had been reached over the
confidentiality claims made. (If work on the Class 2 aspects were to start in advance of such
an agreement, the intellectual property rights to the information contained within the
application would be compromised.)
All workers undertaking Class 3 activities would have to be able to demonstrate their
competence in required techniques before being permitted to register for work on the project.
They must also be approved by Occupational Health (which involves submitting a health
questionnaire, and attending for medical interview. Depending on the project concerned, a
reference sample of serum may be requested: if this is the case, a “leaving sample” would also
normally be requested at the termination of the work on the project.)
___________________________________________________________________
Revised July 05
SG15 Appendix 2 (1)
Specimen project application for higher risk (Class 3) GM project
_________________________________________________________________________________
_
Section 1: General information
All proposers must complete this section
Please tick the box below if any information in this application is to be considered
“Confidential”.
Please identify all such information by marking the relevant section(s) as “confidential.” Note that
“personal information” (Names, addresses, etc) is protected from disclosure under the terms of the Data
Protection Act
√
This form contains confidential information in those sections of the form so marked
1.
Area or Department where the work is
to be carried out.
Please delete as appropriate
2.
Project Supervisor, i.e.,
Member of staff responsible for directing
the work.
Other (please specity): Department of Science
Studies
Clone
Title Dr.
I.M.A.
Initial
Surname
5678
Extension
(c)
Staff involved operationally on the
project, and details of their qualifications/
experience in GM techniques. If
appropriate, give numbers of previous GM
projects.
For undergraduate student classes or
projects, please tick box, and attach
separate lists of participating students and
demonstrators; appropriate lists should be
resubmitted each year the project is run.
NB: only Class 1 activities are permitted
for undergraduate classes.
Name
Position
Qualifications/
experience
Ph.D/ 15 yrs GM
GM
Clone, Dr. I.M.A.
Black, A
Reader in
Science
Ph.D. student
B.Sc. Microbiology/ None
Green, J
Post-doctoral
Ph.D.;10 yrs GM
Brown, Jenny
Post-doctoral
Ph.D. (Zoology); none
White, Carol
Ph.D. student
BSc, Molecular biology; 2
yrs GM
Redditch, Violet
Technician
HNC, Microbiology; 8 yrs
GM
Undergraduate class?
3.
Number / location, and containment level
of laboratory facilities proposed for project.
Please tick box if any of these facilities are
to be used for GM work for the first time.
Room
number/ Containment level
location
101, Science building
2
111A ,
2 (First use)
First use?
√
___________________________________________________________________________
SG15 Appendix 2(2)
Revised July 05
Specimen project application for higher risk (Class 3) GM project
4.
Please tick box and give reference
number if this project is to be considered
as an extension to a previouslyapproved project.
Extension?
Previous Ref. No.
Please then complete appropriate sections in the
remainder of this form, so as to emphasise the
connection between the original project and this
application.
5.
Title of project
Construction of an adenoviral vector with a modified
tissue tropism for possible use in directed therapy of acute
myeloid leukaemia.
6.
Purpose of the project,
stated in layman's terms.
This statement may be
incorporated into the
Project Lay Summary.
The aim of the project is to produce an improved vector for use in
“targeted therapy” of acute myeloid leukaemia. In leukaemia,
abnormal immature lymphocytes are produced in the body, leading
to increased susceptibility to infection, anaemia, tendency to
haemorrhage, etc., and eventually to death. Lymphocytes are not
normally infected by Adenovirus 5, as they lack the appropriate
receptor protein (designated CAR) on the cell surface. The abnormal
lymphocytes have been shown to express a protein (ML1) at very
high levels on the cell surface (Ref: Clone et al, Cell mediated
immunity 45, 123 - 146 (1997)). If this protein could be utilised by a
vector, it would allow the abnormal cells to be targeted by the
vector, and direct the specific introduction of a chemotherapeutic
agent (Diphtheria toxin A fragment) to kill the abnormal cells.
The protein ML1 has also been shown to occur at much lower levels
on the surface of normal lymphocytes, which may therefore be
rendered slightly susceptible to infection by the vector. The protein
has not been detected on any other cell line tested (including human
fibroblasts; HEK cells; HEP-2 cells and various human primary cell
strains.) Scanning of sequence data banks indicates that part of the
ML1 sequence used as the target is only found in primates, and that
no non-human species found in the laboratory environment would
be susceptible to infection.
The project would also examine whether infection of normal
lymphocytes occurs, and if so, whether it could compromise this
approach to treatment of leukaemia.
Confidential information
Revised July 05
SG15 Appendix 2 (3)
Specimen project application for higher risk (Class 3) GM project
_________________________________________________________________________________
_
7.
The scope and expected
results of the project.
(Where relevant: if the
project consists of a
connected programme,
please describe the
expected scope of the
programme.)
The project will initially involve the replacement of part of the
Adenovirus 5 fibre gene with a modified version in which the
recognition site of the sequence will be replaced by a synthetic DNA
sequence. (The fibre protein is the part of the virus which interacts
with the receptor protein on the cell surface.) The introduced
sequence has been calculated to be complementary to (and therefore
bind to) the external portion of the ML-1 protein: studies with
synthetic polypeptides have demonstrated that labelled polypeptide
with this specific sequence preferentially binds to the ML-1 protein
on leukaemic cells. The adenoviral vector to be used is based on a
derivative of Adenovirus 5, which has been deleted for the essential
early genes E1a and E3, and is thus unable to replicate within the
target cells. (The modified virus will be grown/ assembled in the
“packaging” foetal human cell line 293, which expresses the E1a
protein factor required for early transcription in trans.).
At this stage there are no immediate plans to introduce the gene for
the Diphtheria toxin A fragment into the vector, but if the project to
modify the tissue tropism of the vector is successful, this will be the
next stage of this programme. It is expected that the modified
vectors encoding the toxin will comprise the most hazardous
element of this programme. Such vectors would (initially) be
regarded as Class 3 GMMs, and be handled in full Level 3
Containment, until evidence was available concerning their effects
on normal lymphocytes. There are no plans to undertake any clinical
trials of the modified, toxin-inducing vector at this establishment, as
there are no facilities for such work. If the vector appears to be
promising, on the basis of in vitro assessment of its effect on both
normal and abnormal lymphocytes, clinical trials would be
undertaken by our collaborators in the National Leukaemia Trials
Centre in East Grinstead.
Confidential information
___________________________________________________________________________
SG15 Appendix 2(4)
Revised July 05
Specimen project application for higher risk (Class 3) GM project
Section 2: Risk assessment and classification
Please complete
Part A for GMMs, and / or
Part B for GMOs that are not microorganisms.
You will need to complete both parts of this section if your project involves
both GMMs and higher organisms.
[Remember: cell cultures are classified as microorganisms.]
Part A: Projects involving the contained use of genetically
modified microorganisms [“GMMs”].
This section provides you the basic information for the risk assessment form, GM03.
Please give sufficient detail to allow the independent assessor to confirm your risk
assessment. Include all parental / donor and recipient organisms (or indicate the
possible range, identifying the elements of greatest potential risk), and each host-vector
system to be used. Use one entry for each separate system in each heading. Provide
supporting references where necessary.
y
If GMMs are to be imported into the University from elsewhere, the information on the
“identity” of the GMM (from Q1 - Q3) must be obtained from the prospective supplier.
[N.B.: it is permissible to use a risk assessment prepared by the supplier, provided that
this gives the information required in the headings below.] Such GMMs must NOT be
obtained before obtaining project approval.
Please tick box if the GMO was constructed elsewhere and
Â
is being brought onto University premises
1.
The identity, source and function of
each sequence of genetic material to
be inserted.
Identify all potentially harmful
characters such as toxin genes, or
other pathogenic functions. [Please
extend each section as necessary to include
all donor or parental organisms, etc. - use
one heading for each insert.] State ACDP
or other hazard categorisation [e.g., MAFF]
of any donor or parental microorganism.
Where relevant, identify: any antibiotic
resistance markers and potential for
transmission; mutations attenuating
virulence and possibility of reversion;
involvement in environmental processes;
natural habitat and geographic distribution.
Revised July 05
a) Synthetic sequence, calculated to produce a protein binding site
complementary to human leukaemia ML-1 protein. (This
protein expressed to high levels on surface of abnormal
lymphocytes and stem cells in acute myeloid leukaemia; only
detected at very low levels on surface of normal cells.)
B) Diphtheria toxin A subunit gene - synthetic DNA sequence
derived from Corynebacterium diphtheriae (ACDP category
2). The A subunit is responsible for the toxic effects of
Diphtheria toxin, which has been show to bind to and inhibit
the elongation factor EF-2 in eukaryotic protein synthesis.
Confidential information
SG15 Appendix 2 (5)
Specimen project application for higher risk (Class 3) GM project
_________________________________________________________________________________
_
2.
State identity [Species, strain(s)]
and ACDP hazard category of all
recipient microorganisms
[See ACGM Compendium of Guidance for
information on hazard categories of disabled
bacterial host strains.] For cell cultures, give
strain/ line information as well as species. In
all cases, identify any potential harmful
effects of the unmodified recipient, including
the possibility of pathogenic agents present
in the culture and possible effects they could
cause (e.g., pathogenic viruses in cell
cultures recombining with a viral vector.)
Use one line for all strains of each species of
recipient (with the same level of disability.)
3.
Identity of the vector(s), and nature
of any potential harmful properties.
If the vector is a plasmid or prokaryotic virus,
state whether it is mobilisable, and/or
capable of being transferred to another host.
For vectors derived from eukaryotic viruses,
state hazard class of parent virus. In all
cases, identify circumstances in which the
vector can replicate, or is maintained in the
recipient. Where vectors are not used,
identify the means of insertion of DNA into
the recipient (e.g., use of the "gene gun".)
Adenovirus 5, strain deltae1e3: based on ad-5 (=ACDP
cat. 2), with deletion of early genes e1 and e3.
Replication negative, assessed as ACDP 1 (similar
strains used in human gene therapy: ref. Smith and Jones,
PNAS, 231, 1234 - 1243 (1997)
B) Human Leukaemic cell line, strain ml-1, derived from
patient isolate (target cells: will be “recipient” of
modified vector.)
C) Human cell line 293, expressing adenovirus gene e1a,
used for replication and assembly of modified
adenovirus.
D) Human normal lymphocyte line (“control target”)
A).
ACDP recommend that all human-derived cell lines be
regarded as hazard category 2. All lines to be used have
been shown to be free of adventitious viruses.
Confidential information (b)
a) Intention is to produce adenoviral vector
containing a modified fibre gene, such that the
vector will adsorb to the ml-1 protein and
thereby infect cells expressing this protein.
Synthetic sequence to be inserted into isolated
ad5deltae1e3 DNA as a replacement for
sequence in the fibre gene, which codes for
receptor recognition and so determines the
tropism of the virus. (full sequence of this strain
of adenovirus 5 DNA is known.) Ligation
techniques will be used to assemble the
recombinant adenoviral genome, followed by
transfection into 293 packaging cell line.
In this instance, the “target” of the modification
is the vector, adeno5 deltae1e3 : see 2(a) above.
Vector cannot express any genes in the absence
of the e1a gene, nor can it replicate its DNA,
(and is thus eventually eliminated from its target
cells if they survive the infection.) Vector is
efficiently packaged in a selected strain of 293
cells (human embryo kidney derived packaging
cells, containing tandem-integrated e1a genes.
These are expressed to a high level, allowing
transcription, replication and packaging of
adeno5.) (ACDP recommend that all humanderived cell lines be regarded as hazard category
2; in this instance, the presence of the adenovirus
e1a gene would not increase the hazard of the
cells above hazard category 2.)
(Continued on next page)
___________________________________________________________________________
SG15 Appendix 2(6)
Revised July 05
Specimen project application for higher risk (Class 3) GM project
3. (Continued)
4.
Natural host (if any) of recipient
organism(s) and routes of
transmission/infection (if known).
Include known or suspected
adventitious viruses in cell
cultures.
Revised July 05
When modified (see 1a), the vector is expected to
acquire the ability to infect the abnormal
lymphocytes, but the presence of low levels of
ML-1 protein on the surface of normal cells may
provide a route of infection of normal
lymphocytes and their parent stem cells. As these
are not normally infected by adenoviruses, it is not
possible to say whether their infection by a nonreplicating adenovirus would kill the cells due to
possible cytopathic effects of adenoviral gene
expression. As expression of the “late” (structural)
genes of adenovirus only occurs after DNA
replication, there is expected to be little or no gene
expression from the Adeno5 DeltaE1E3 modified
for the adenovirus fibre gene. Until appropriate
evidence has been obtained to justify downgrading
the hazard to Category 1, such viruses will be
regarded as Hazard Category 2.
B) When the gene for the toxin a subunit gene
(question 1b) is also inserted, it will be used to
replace the e1a gene, under the control of the e1a
promoter, and thus be expressed in the cells
following infection. (the continued absence of the
e1a gene prevents any dna replication or
expression of any of the other adenovirus genes
remaining in the vector.) It is therefore expected
that the target cells would be killed following
infection, and the possibility exists that a
proportion of normal lymphocytes could also be
killed by the modified vector. In the 293
packaging cell line, expression of the toxin gene
during early transcription is expected to reduce the
yield of virus by approximately 2 logs compared
with the vector lacking this gene.
Confidential information (a) and (b)
a) Humans: wild-type Adeno5 infects cells in the
respiratory tract, transmitted by droplet inhalation.
The “parental vector” strain DeltaE1E3 also
infects the same types of cells by the same route.
b) None
c) None
d) None
SG15 Appendix 2 (7)
Specimen project application for higher risk (Class 3) GM project
_________________________________________________________________________________
_
5.
For each GMM, state: Likelihood of
harmful effects being produced as a
result of infection/ colonisation of
humans and/ or the environment;
likelihood of "insert" product [protein or
nucleic acid] being produced/ secreted
in an active form by the GMM (based on
the expected levels of expression);
targets and potential effects of each
product. For “antisense” RNA
transcripts, identify function of gene(s)
interfered with and indicate possible
effects on the resultant GMO. Take into
account any possible need for an
inducer to initiate protein production. For
genetically modified viruses or viral
vectors, include possibility of
recombination with unmodified or related
viruses infecting the same target cell.
Susceptibility of normal lymphocytes/ stem cells
expected to be low, due to very low levels of ML1 protein on cell surface. The modified vector is
unable to replicate in the target (leukaemic) cells,
but expression of the residual adenovirus genes is
likely to disrupt cellular metabolism and may
cause cell death. Vector containing the modified
fibre gene is expected to target the ML-1 protein
on abnormal lymphocytes, and render them
susceptible to infection by the vector. The vector
is unable to replicate or express any genes within
the target cells because of the lack of the essential
E1a early gene function, and thus the only effect
on the cells could be a by-product of the infection
process disrupting normal cellular metabolism.
Such effects would be limited to the cells actually
infected by the vector. Because of the presence of
low numbers of the ML-1 molecule on the surface
of normal lymphocytes, it is possible that they
could also be rendered susceptible to infection, but
the effect of any possible disruption is likely to be
much less.
Vector containing the gene for the Diphtheria
toxin A subunit is expected to kill those cells that
it infects, but the lack of the E1a gene will prevent
expression of any viral genes, or viral DNA
replication. Death of the cell would prevent the
maintenance, transfer or transmission of the
modified DNA to a subsequent population of
target cells, and so the genes could not be
transmitted to other organisms in the environment.
If this part of the programme is successful,
subsequent insertion and expression of the gene
for the diphtheria toxin A subunit is expected to
result in the death of the target cells.
Harmful effects on normal immunocompetent
humans regarded as unlikely, and limited to cells
initially infected by the modified virus. Change in
tropism of the modified virus will preclude
possibility of joint infection by wild-type
adenovirus (which targets cells in the respiratory
tract) and the modified virus.
Confidential information (a), (b) and (c)
___________________________________________________________________________
SG15 Appendix 2(8)
Revised July 05
Specimen project application for higher risk (Class 3) GM project
6.
Severity of all potential harmful
effects of the GMM. (Take into account
“insertional” effects; expression of the
insert and/or genes comprising the
vector, vector replication; stability/
maintenance of the vector in the
recipient; potential for transmission of
genes to other organisms, survival in the
environment, etc.) Do not forget
“marker” genes such as antibiotic
resistance genes and the possibility of
compromising antibiotic treatment.
7.
State maximum culture volume to be
used.
8.
Identify all possible routes of
exposure to the GMM.
9.
Potential for exposure (this will
depend on the volume of culture and the
type of operation; for example, potential
for aerosol generation could be “high” for
centrifuging operations.] .
10.
If potential for exposure is “high” or
“medium”, state cause of potential
exposure, and give details of measures
to be used to protect both humans and
the environment.
Revised July 05
Any potential harmful effects likely to be mild and
transient, as far as normal cells are concerned: in
the "worst case", could cause temporary depression
of the immune system.
No effects expected on organisms other than
humans.
Maximum culture volume <50 ml.
Predominantly by aerosol route: Procedures to be used
to minimise aerosol generation: all work to be
performed in microbiological safety cabinet or
apparatus offering a similar degree of containment.
Sealed rotors to be used for centrifuging operations.
Gloves to be worn to prevent exposure by skin contact.
Low
N/A
SG15 Appendix 2 (9)
Specimen project application for higher risk (Class 3) GM project
_________________________________________________________________________________
_
11.
State methods to be used for
decontamination of waste and
equipment. If disinfection to be used,
identify disinfectant and state conditions
of use. State procedures used to monitor
effectiveness of disinfection.
A) Class 2 work: All solid waste to be autoclaved
(135o for 15 minutes) before discard to skip;
contaminated recyclable labware (glass; plastic) to
be decontaminated by soaking in freshly-prepared
1% Virkon for 1 hour before collection for
autoclaving and washing. Liquid waste (e.g.,
culture supernatants) collected into fresh 1%
Virkon and then autoclaved as above before
discard to drain. Surfaces to be swabbed with
solution of sodium dichloroisocyanurate (200-ppm
available chlorine) after work finished. Spills of
infectious material to be treated by covering with
Virkon powder and leaving for minimum of 20
minutes.
b) If/ when Class 3 activity started, waste for
discard will be collected in autoclave bins, and
fitted with leak-proof lids for transport to
autoclave. Lids to be loosened to allow steam
penetration on loading into autoclave, immediately
prior to autoclave operation. “Recyclable” labware
to be disinfected in CL3 facility before loading into
autoclave bins as for “discard” material. Other
disinfection procedures as for CL2 work described
in (a).
All disinfection procedures shown to cause at least
4 log. reduction in infectivity titre of wild-type
Adeno5 with freshly-prepared disinfectants.
___________________________________________________________________________
SG15 Appendix 2(10)
Revised July 05
Specimen project application for higher risk (Class 3) GM project
A2: Risk assessment.
The risk assessment is based on the information on possible harmful effects, and the
potential for exposure given in Section A1.
12.
13.
Provisional hazard classification of
the genetically modified microorganism[s] (i.e., based on hazard
classification of the recipient organism,
as modified by the insertion / expression
of the vector or foreign DNA.)[Use one
line for each species of GMM, quoting all
strains of with the same hazard class.]
Environment likely to be exposed
to the GMM(s)
14.
Possible risks to (a) human health
and (b) the environment: On the basis
of the provisional hazard level allocated
to the GMM(s), identify all possible risks
to human health and to the environment.
15.
Provisional containment level required
to control the risk to (a) human health
and (b) to the environment.
a) Vector modified for tissue tropism assessed as
hazard category 2
Vector modified for expression of DipA toxin
gene, hazard category 3, based on possibility of
infection and killing of normal lymphocytes.
Confidential information
a) and b): Laboratory environment only: Containment
procedures to be adopted will minimise escape to the
wider environment.
Risks to human health only: Adenovirus 5 is
specific to humans. Although the aim is to modify
the tissue tropism of the virus, the modification
has been directed against a human protein on the
surface of leukaemic cells, and it is not expected
that there will be any gross changes that would
allow infection of cells from other species or
human cell types. This aspect will be checked by
testing the modified vector against a range of cell
cultures of human, mammalian and avian origin.
The risks to human health arise mainly because of
the possibility of infection of normal lymphocytes
expressing low levels of the ML-1 protein. Even
though the vector cannot replicate in such cells, it
may cause cytopathic effects, and therefore
warrants containment level 2, until such time as
evidence is obtained to justify a lower
containment level.
The vector with the inserted toxin gene could also
kill normal lymphocytes, and thereby warrants a
higher degree of containment than (a). Such
infection might cause serious illness in infected
individuals, especially anyone suffering from
immunosuppression or under immunosuppressive
therapy.
Confidential information
a) Containment level 3
b) N/A
Revised July 05
SG15 Appendix 2 (11)
Specimen project application for higher risk (Class 3) GM project
_________________________________________________________________________________
_
16.
17.
18.
Can all the risks to human health and
the environment be adequately
controlled by standard measures
appropriate to the provisional
containment level? (See Tables 1a - 1c
in Schedule 8 to the Regulations)
Yes
If the answer to Q16 was “No”,
specify risks not controlled by standard
measures for the provisional
containment level, and identify any
special arrangements required to
manage this risk. Otherwise, go to Q18.
N/A
Final Containment level required to
control the highest risks associated with
the project [this is numerically equal to
the “Class of Activity”][See Note 6 in
“Notes for Guidance” on p2 of this form]
Please delete as appropriate
Containment level 3 ⇒ Class 3 Activity
___________________________________________________________________________
SG15 Appendix 2(12)
Revised July 05
Specimen project application for higher risk (Class 3) GM project
Statement on the overall risks associated with this project.
Use the information provided in Parts A and/ or B of Section 2 to draft this statement. You
should identify the most hazardous elements of the project, and any particular procedures
necessary to control the associated risks. Extend the section as necessary.
It is expected that the most hazardous elements of this proposed programme of work
will arise when the Diphtheria toxin gene is inserted into the modified Adenoviral
vector. It is expected that expression of the inserted gene in the target cells will result in
the death of those cells. In patients with leukaemia, the intention is to target the specific
destruction of cells over-expressing the ML-1 protein. However, there is a possibility
that the level of expression of the target ML-1 protein in normal lymphocytes could
result in some of these cells also being rendered susceptible to infection by the modified
vector. Although the vector will be unable to replicate in the cells that it infects,
infection could result in the death of all target cells.
For the vector with modified tropism alone, it is expected that the risks will be lower,
and unlikely to be greater than the parent adenovirus, since any effects of expression of
the vector in the cells will be limited to early events in the “replication cycle” of the
vector (such events may cause disruption to cellular metabolism in the target cells, but
as the vector is unable to replicate, those effects will be limited to the cells actually
infected by the vector.) Because of the possibility that normal lymphocytes could be
infected and killed by the vector with modified tropism, there is a possibility that
accidental infection of workers could cause a temporary depression of the immune
system. Care will be taken to exclude any workers who may be more at risk (for
example, those under immunosuppressive treatment or having a compromised immune
system). The rate of turnover of normal lymphocytes is such that any normal cells
killed would be expected to be replaced within a week of infection.
Protection of workers will be achieved by adopting standard Containment level 2
procedures for work with the vector targeted towards the ML-1 protein, and
Containment level 3 for work with the vector containing the inserted Diphtheria toxin A
subunit gene. The waste control procedures to be adopted will prevent the modified
virus escaping from the laboratory. Even if an escape did occur, it is extremely unlikely
that this would have any wider environmental impact, as the modified virus is unable to
replicate (and thus cannot persist in the environment) and is not expected to be able to
infect cells of species other than humans. This is an important aspect of any potential
use of the modified virus in the treatment of leukaemia in human patients, as they would
not be confined or segregated from the community whilst undergoing treatment. The
adoption of Containment level 3 is regarded as a necessary precaution until evidence to
warrant downgrading the containment level is accumulated. Clinical trials of any agent
produced as a result of this programme would not be planned or proposed unless and
until it could be shown that the modified vector viruses were safe for such use.
Revised July 05
SG15 Appendix 2 (13)
Specimen project application for higher risk (Class 3) GM project
_________________________________________________________________________________
_
Section 3: Declarations and lay summary of the project.
[If the form is completed by computer and returned by e-mail, this and the following page should be printed off, signed, and
returned by internal mail to the Safety Office marked “For the attention of SCBS”.]
1. Proposers' Declaration:
I am submitting this application for approval at the next meeting of the Safety Subcommittee for
Genetic Modification. I declare that work will not commence on this project until SCBS has given its
approval, and all workers will be properly registered with the Occupational Health Medical Adviser.
Name (BLOCK LETTERS) ........IAN ARTHUR MICHAEL CLONE............
Signed
................................................Date .......31 August 2001.......
2. Confidentiality Statement. [Please delete if not applicable.]
I wish to claim the information given in sections One - (7) & (8); Two - A1(1), (2), (3) & (5); A2 (12) &
(14)……………………of this form as “Confidential”.
This information is given on the understanding that it is only received by persons properly
authorised by the Safety Sub-Committee for Genetic Modification, who have signed a
declaration regarding the non-disclosure of such confidential information.
Signed
......................................................................................Date ..........................................
3. Supporting Declaration
(The person supporting this proposal must not be involved in the project being proposed.)
I support the presentation of this proposal to the subcommittee.
Name (BLOCK LETTERS)
....................................................................……........................................
Signed
............................................Date ..........................................
Status in organisation:
Head of Department
(please delete as necessary)
PLEASE CHECK THAT:
• ALL RELEVANT SECTIONS OF THIS FORM HAVE BEEN COMPLETED,
• REQUESTED SUPPORTING INFORMATION IS SUPPLIED, AND
• DECLARATIONS ARE SIGNED.
Proposers must deposit a copy of this completed, signed form with the Health
& Safety Co-ordinator/ Health & Safety Technical Administrator, as well as
keeping a copy for their own records. Please Note:
• Proposals giving insufficient information may be subject to delay if the forms are returned for the
proposer to supply more information.
• For all Class 2 (or higher) projects, Proposers will be invited to present their case to a technical
sub-group of SCBS for discussion and assessment prior to a full SCBS meeting.
___________________________________________________________________________
SG15 Appendix 2(14)
Revised July 05
Specimen project application for higher risk (Class 3) GM project
Lay summary of the project
This statement is the main source of information for "lay" members of the Committee. It will
be reproduced and submitted to the Committee for formal approval. Sufficient detail, in nontechnical terms, should therefore be given to allow them to understand the aims of the
project, and to assess the associated risks. For higher risk projects, this summary will be
accompanied by a report from the Technical Assessment group.
Outline the aims and expected results of the project. Where possible, avoid the use of acronyms and
abbreviations, and explain or spell out any used. ("DNA" and "RNA" need not be spelt out.) Identify the main
hazards and risks both to humans and to the environment. Describe the measures to be put in place to control
the identified risks. Clearly identify the class of the activity for all proposals for work with genetically modified
microorganisms.
[Expand box as required.]
Leukaemia is a common cancer of the blood cells, with about 5000 new cases being diagnosed each year in
the UK. This project aims to provide a basis for improving the therapy to combat acute myeloid leukaemia.
(This type of leukaemia accounts for about 80% of all new cases, and normally has a low post-6 month
survival rate). We have previously shown that the abnormal lymphocytes (white blood cells) present in this
form of leukaemia have a high level of a characteristic protein on their surface. By contrast, normal
lymphocytes have very low levels of this protein on their cell surface, typically about 1 molecule per cell.
Other normal human cells either do not have this protein, or similarly have very low levels. The intention is to
use this protein as a "cellular target" for a defective form of a human virus, which will then be used to
"deliver" a toxin to specifically kill the abnormal cells. (The virus to be used is an adenovirus, which normally
causes one type of the common cold.) The defective virus cannot replicate in the cells it infects and so has to
be produced in the laboratory in special "helper cells". Because the virus can no longer replicate, it can no
longer cause disease, and only requires containment level 1.
The first phase of the project is to modify the adenovirus so that it will specifically recognise the abnormal
lymphocytes, instead of the cells in the respiratory tract that it would normally infect. The main risk to
humans in good health is that the virus may also infect normal cells because of the presence (albeit at a very
low level) of the target protein on the surface. An infection would be limited to the initial cell infected,
because the virus cannot replicate. Such an infection might cause a temporary depression of the immune
system if normal lymphocytes are affected by the infection. This stage will be undertaken at Containment
level 2, and the possible interaction and targeting of normal lymphocytes will be thoroughly examined before
any attempts are made to commence the second phase of the project.
The second phase aims to insert a piece of synthetic DNA into the adenovirus, which will be used as a
delivery vehicle (vector) to introduce this synthetic gene into the leukaemic cells. The synthetic gene is
identical in sequence with part of the Diphtheria toxin, thus infection of the abnormal lymphocyte should
result in the diphtheria toxin being produced within the cell. When the toxin is synthesised, it stops protein
synthesis in that cell, and so kills the cell. The net effect is that the abnormal lymphocytes are specifically
killed, whilst the non-leukaemic cells survive.
Construction and use of the adenovirus with the inserted Diphtheria toxin gene will initially be treated as a
Class 3 activity because it has the potential to introduce a cellular toxin into cells and kill them. This
precautionary approach may produce evidence that will warrant downgrading the activity to Class 1 to
eventually allow use of the modified virus in clinical trials on patients (which will be a different project to be
conducted elsewhere). There are no risks to the environment because the virus to be used is derived from a
human-specific pathogen and will not infect cells of other species. This specificity is not expected to change
following modification to recognise the protein found on the surface of the leukaemic cells.
Reminder:
• All workers named in this proposal must also complete and return a worker registration form [ref.
GM02f] to the Occupational Health Medical Adviser.
• The worker registration form(s) should only be submitted after the Project Supervisor has been
notified in writing that the project has been approved.
• Worker Approval is subject to any conditions imposed by SCBS when the project is approved. An
individual may not start work on the project until written confirmation has been received stating that
his/her registration has been approved.
Revised July 05
SG15 Appendix 2 (15)