microRNA Research
Lenti-miR microRNA
Precursor Clones
™
Overexpress Individual microRNAs or Pooled Virus Library
Precursor Clone Collection
Highlights
System Biosciences (SBI) has the largest
Native microRNA
hairpin structure
retained.
commercially-available collection of
microRNAs that are cloned into lentiviral
vectors. SBI’s constructs can be stably
• Largest collection of microRNA
precursor clones available in
lentivectors
SBI’s microRNA
precursor clones
are the real thing.
expressed in a wide variety of cell types,
• Native microRNA context ensures
accurate and robust mature
microRNA production
3’
5’
as opposed to synthetic microRNAs that
of the native stem loop structure and 200-400 base pairs of upstream and downstream flanking genomic
• Confirm positive expressing
cells with copGFP for convenient
sorting of transfected or
transduced cells
sequence. This unique feature ensures that the microRNAs expressed from SBI’s constructs will be correctly
• Easily create stable cell lines
processed in the cell into mature microRNAs.
• High-throughput microRNA
phenotypic screens using pooled
Lenti-miR virus library
can only be transiently tested in cells.
Native genomic sequences flanking hairpin (~400 bp)
Each construct in SBI’s collection consists
MCF-7 Breast Cancer Cells
Transduced with Lenti-miR-205
Phase Contrast
GFP Fluorescence
RSV 5’LTR
The microRNA precursors
expressed from SBI’s
Lenti-miR vectors produce
mature microRNAs via
the host cell’s processing
machinery.
Ψ
AmpR
cPPT
pUC ORI
CMV
Lentiviral Pre-miR-205 transduction
SV40 ORI
SV40 poly-A
3’∆LTR
Mature miRNA Expression
MCF-7 Cells infected (MOI 5:1)
with Lentivirus made from
PMIRH205PA-1 (pre-miR-205)
construct. Images taken after
48 hours. Expression of mature
miR-205 was validated and
quantitated using qPCR assays.
12
Relative Expression Level
(Normalized to U6)
10
8
6
4
3’
dsRNA
Intermediate
microRNA
Precursors
copGFP
WPRE
Drosha
processing
Dicer
processing
5’
3’
5’
3’
5’
miR : miR*
2
0
EF1
Dicer
miR-205
Control
3’
5’
miR* separation
www.systembio.com/lentimir
5’
AGO2
3’
miRISC complexes
Lenti-miR™ microRNA Precursor Clones
Screen All microRNAs Simultaneously for Unique
Phenotypes
SBI’s Lenti-miR virus library contains the pooled Lenti-miR precursor
clone collection in a ready-to-infect lentiviral preparation. The library
is designed for studying phenotypic effects associated with the over-
Pooled Lentiviral microRNA Screens
Transduction
with Lenti-miR
microRNA Library
expression of individual microRNAs in hard-to-transfect mammalian
Infected
Target Cells
cell lines, primary cells, non-dividing cells and even whole animal
studies. The lentivirus preparation is pseudotyped with VSV-G for
broad cellular tropism. The microRNA or microRNAs responsible for
generating the phenotypes of interest may be recovered through
Treatment
to induce
phenotype
Treated Cells
simple genomic PCR using lentivector-specific primers followed by
direct sequencing of microRNA precursor clones recovered.
Perform One Transduction and Easily Identify the
microRNAs Involved in Your Phenotypic Screen
Each virus within the pool will express an individual microRNA
precursor in its native context while preserving hairpin structures
to ensure biologically relevant interactions with endogenous
processing machinery and regulatory partners.
Lentivirus Infection QS Data
Tumor Invasion
Cell Differentiation
Apoptosis
Amplification
of microRNA
effectors from
selected cells
Identification of microRNA Effectors
Simple Genomic PCR to
Identify microRNA Effectors
HT1080 Cells
Phase Contrast
HEK 293 Cells
Selection
for phenotype
CMV
MicroRNA
Precursor clone
5’ LVL Primer
copGFP
3’ LVL Primer
MicroRNAs
M
16 29a 210 155 651 203 34a 125b
(bp)
GFP Fluorescence
1000
500
Pooled Lentivirus Particles of all SBI’s MicroRNA Precursors
The most comprehensive microRNA Precursor
Clone Collection in lentiviral vectors available
anywhere.
Product Description
Catalog #
Size
MicroRNA Precursor Pooled Library
(Pre-packaged) >10^7 IFUs/aliquot
PMIRHPLVA-1
4 virus aliquots
MicroRNA Precursor Pooled Library
(HT, Pre-packaged) >10^7 IFUs/aliquot
PMIRHPLVAHT-1
10 virus aliquots
We Also Offer Custom Services
System Biosciences offers a wide-range of custom services to support your research, allowing you to spend less time making tools, and more
time making discoveries. To learn more, visit our website at www.systembio.com/service or call us at 888-266-5066.
System Biosciences, Inc.
1616 North Shoreline Blvd.
Mountain View, CA 94043
Toll Free: 888.266.5066
Fax: 650-968-2277
Email: [email protected]
www.systembio.com
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