DreamFect TM Gold Transfection Reagent - Results
OZ Biosciences is delighted to announce the launching of a new lipid-based transfection reagent, DreamFect TM
Gold. This formulation is based on OZ Biosciences Triggered Endosomal Escape Technology (TEE-Technology).
It was specifically developed to achieve high transfection efficiency (percentage of cells transfected) combine
with superior transgene expression level due to its improved cytoplasmic release process and complete
biodegradability. In this way, DreamFect Gold is the ideal transfection reagent in a wide variety of cells.
Main
1.
2.
3.
4.
5.
6.
DreamFect TM Gold features are:
Highly efficient & Biodegradable
Achieve superior transgene expression level than any other reagents
Multipurpose (various types of nucleic acids)
Universal (cell lines and primary cells)
Simple, Ready-to-use and Rapid
Serum compatible
Nucleic acid types
DreamFect TM Gold Transfection Reagent is suitable for all type of nucleic acids including: plasmid DNA, siRNA,
oligonucleotides, linearized DNA, double stranded RNA, mRNA, shRNA.
Cell types
DreamFect
TM
Gold is suitable for numerous cells. This reagent has been successfully tested on several cell
lines (see Table 1, page 2). If a particular cell type is not listed, this does not imply that DreamFect TM Gold is
not going to work. OZ Biosciences is maintaining an updated list of cells successfully tested that is available on
the website: www.ozbiosciences.com. You can also submit your data to [email protected] so we can
update this list and give you all the support you need.
DreamFect TM Gold transfection efficiency on various cell lines
DreamFect Gold Transfection Efficiency - I
90
80
% Transfected Cells
70
60
50
40
30
20
10
0
A549
BEAS-2B
BHK21
COS-7
CHO
HELA
NIH3T3
Cell Line
OZ Biosciences – DreamFect ™ Gold Transfection Reagent - Results – v1.2
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DreamFect Gold Transfection Efficiency - II
60
% Transfected Cells
50
40
30
20
10
0
MDCK
Jurkat
K562
L929
U87
VERO
SHSY5Y
HaCat
Cell Line
Several cell lines (1x105 cells/ well) were transfected with 1 µg of pEGFP plasmid DNA per well in a 24-well plate.
Transfections were performed with 4 µL per well of DreamFect Gold transfection reagent. Percentage of
transfected cells were measured 24h post transfection by cytofluorimetry.
Table 1: Example of cells successfully transfected with DreamFect TM Gold Transfection Reagent.
Cell Lines
Cell Type
293, 293T
3T6
A549
B16-F10
BEAS-2B
BHK-21
CHO, CHO-K1
COS-1 , COS-7
CV-1
H441
HaCat
HEK293
HeLa, HeLa-S3
Jurkat *
K562*
L929
MCF-7
MDCK
N2A
NIH-3T3
PC-12
RAW
SH-SY5Y
U87
Vero
Kidney
Embryonic Fibroblasts
Non-small cell lung carcinoma
Melanoma
Bronchial Epithelial
Kidney
Ovary (epithelial like)
Kidney
Fibroblast-like (Kidney)
Lung epithelial
Keratinocyte
Kidney
Cervix carcinoma
Acute T cell lymphocyte
Myelogenous leukemia
Fibrosarcoma cells
Breast adenocarcinoma
Kidney
Neuroblastoma
Fibroblasts
Pheochromocytoma (Adrenal)
Macrophage
Neuroblastoma
Glioma
Kidney
Species
% transfected
cells
Transgene
expression level
Human
Mouse
Human
Mouse
Human
Hamster
Chinese Hamster
Green Monkey
Monkey
Human
Human
Human
Human
Human
Human
Mouse
Human
Dog
Mouse
Mouse
Rat
Mouse
Human
Human
Green Monkey
85-95 %
N.D.
80-90 %
N.D.
65-75 %
65-75 %
80-90 %
65-75 %
N.D.
30%
15-25 %
85-95 %
80-90 %
20-30 %
45-55 %
20-25 %
30-35 %
55-65 %
70-80 %
60-90 %
N.D.
10-20 %
35-45 %
40-50 %
50-55 %
+++
N.D.
+++
N.D.
++
++
+++
+++
N.D.
N.D.
+
+++
+++
+
++
+
N.D.
++
+++
++
N.D.
+
+
++
+++
OZ Biosciences – DreamFect ™ Gold Transfection Reagent - Results – v1.2
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Species
Primary Cells
% transfected
cells
Transgene
expression level
N.D.
N.D.
N.D.
N.D.
Glioma
Human
Hippocampal Neurons
Rat
* Suspension cells
ND: Not determined
+++:
≥ 40 ng transgene expression level in a well of 96-well plate
++:
20 to 40 ng transgene expression level in a well of 96-well plate
+:
≤ 20 ng transgene expression level in a well of 96-well plate
GFP expression in different cells transfected with DreamFect TM Gold
CHO-K1
HeLa-S3
BEAS-2B
MDCK
Jurkat
Primary Hippocampal Neurons
OZ Biosciences – DreamFect ™ Gold Transfection Reagent - Results – v1.2
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HEK293
Several cell lines (1x105 cells/ well) were transfected with 1 µg of pEGFP plasmid DNA and 4 µL of DreamFect
Gold reagent per well in a 24-well plate. EGFP expression was monitored 24 h after transfection by fluorescence
microscopy except for Neurons which were analyzed 72h post-transfection.
β-Galactosidase expression in different cell lines transfected with DreamFect TM Gold
BHK-21
MDCK
HeLa-S3
NIH-3T3
Various cell lines (1x105 cells/ well) were transfected with 1 µg of pLacZ plasmid DNA and 4 µL of DreamFect
Gold reagent per well in a 24-well plate. β-Galactosidase expression was revealed 24 h post-transfection using
OZ Biosciences’ X-Gal staining kit (catalog # GX-10003).
OZ Biosciences – DreamFect ™ Gold Transfection Reagent - Results – v1.2
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Superior transgene expression level with DreamFect TM Gold
Relative Amount of β-galactosidase
Higher Protein Expression Level with DreamFect Gold in common cell lines
350
Reagent X
DreamFect Gold
300
250
200
150
100
50
0
HeLa
COS-7
CHO
A549
NIH-3T3
BEAS-2B
BHK21
Cell Line
Various cell lines (15x103 cell / well) were transfected with 0.25 µg of pLacZ plasmid and 1 µL of DreamFect
Gold reagent per well in a 96-well plate. β-Galactosidase expression was monitored 24 h after transfection using
OZ Biosciences’ ONPG β-galactosidase assay kit (catalog # G010001). Quantity of β-galactosidase in each well
was measured and results are expressed as relative values.
Relative Amount of β-galactosidase
Higher Protein Expression Level with DreamFect Gold in hard-to-transfect cell lines
350
Reagent G
DreamFect Gold
300
250
200
150
100
50
0
VERO
K562
U87
MDCK
Cell Line
Various cell lines (15x103 cell / well) were transfected with 0.5 µg of pLacZ plasmid and 2 µL of DreamFect Gold
reagent per well in a 96-well plate. β-Galactosidase expression was monitored 24 h after transfection using OZ
Biosciences’ ONPG β-galactosidase assay kit (catalog # G010001). Quantity of β-galactosidase in each well was
measured and results are expressed as relative values.
OZ Biosciences – DreamFect ™ Gold Transfection Reagent - Results – v1.2
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DreamFect Gold outperform other transfection reagents
Comparison with other transfection reagents - I
Reagent D
180
DreamFect Gold
Relative Amount GFP
160
Reagent L2
Reagent F
140
120
100
80
60
40
20
0
NIH3T3
HeLa
CHO
Cell Line
5
Cell lines (1x10 cells / well) were transfected with 1 µg of pEGFP plasmid DNA and 4 µL of DreamFect Gold
reagent per well in a 24-well plate. DreamFect™ Gold transfections were performed as described in the
instruction manual with 4 µL of reagent / µg of DNA. The other transfection reagents were assayed according to
the manufacturer’s instruction. The average intensity of GFP was measured in each well by cytofluorimetry and
results are expressed as relative values.
Comparison with other transfection reagents - II
250
DreamFect Gold
Reagent D
Realtive Amount of β-galactosidase
Reagent G
Reagent LP
200
Reagent F
150
100
50
0
NIH3T3
MDCK
SH5YSY
Several cell lines (75x103 cells / well) were transfected with 0.5 µg of pLacZ plasmid DNA and 2 µL of DreamFect
Gold per well in a 96-well plate. The other transfection reagents were assayed according to the manufacturer’s
instruction. β-galactosidase expression was measured 24 h post-transfection using OZ Biosciences’ ONPG βgalactosidase assay kit (catalog # G010001).
OZ Biosciences – DreamFect ™ Gold Transfection Reagent - Results – v1.2
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DreamFect Gold is non toxic
Effect of various transfection reagents on cell viability
Total Amount of Protein (µg)
90
80
70
60
50
40
30
20
10
0
Non
transfected
DreamFect
Gold
Reagent IB
Reagent G
Reagent LP
Reagent LX
HeLa cells (75x103 / well) were transfected with 0.5 µg of pLacZ plasmid and 2 µL of each reagent per well in a
96-well plate. The total amount of protein per well was monitored 24 h after transfection using OZ Biosciences’
Bradford Protein Assay kit (catalog number BA00100).
DreamFect Gold is suitable for suspension cells
DreamFect Gold efficiency (protein expression) on
suspension cells
DreamFect Gold efficiency (%) on suspension cells
300
45%
DreamFect
40%
DreamFect Gold
Relative amount of β-Galactosidase
% GFP expressing cell
50%
35%
30%
25%
20%
15%
10%
5%
0%
Jurkat
K562
DreamFect
250
DreamFect Gold
200
150
100
50
0
Jurkat
K562
Cell lines (2 x105 cells / well) were transfected with 2 µg of pEGFP or pLacZ plasmid DNA and 6 µL of DreamFect
Gold or 4 µL of DreamFect per well in a 24-well plate described in the instruction manual; Cells were analyzed
24 hours post-transfection by flow cytometry for percentage of GFP expressing cells or by using OZ Biosciences’
CPRG β-galactosidase assay kit (catalog # G010002).
OZ Biosciences – DreamFect ™ Gold Transfection Reagent - Results – v1.2
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DreamFect and DreamFect Gold comparison study
•
DreamFect & DreamFect Gold have comparable efficiency in term of % of transfected cells
Transfection Efficiency of DreamFect Gold vs DreamFect
DreamFect Gold
DreamFect
90
80
% Transfected Cells
70
60
50
40
30
20
10
0
•
HeLa
COS-7
NIH3T3
K562
MDCK
DreamFect Gold provides higher transgene expression level than DreamFect due to improved
cytoplasmic release mechanism and better biodegradability.
Higher Protein Expression Level with DreamFect Gold vs DreamFect
Relative Amount of β-galactosidase
400
DreamFect
DreamFect Gold
350
300
250
200
150
100
50
0
HeLa
COS-7
NIH-3T3
K562
MDCK
Higher Protein Expression Level with DreamFect Gold
Relative Unit of Luciferase
140
DreamFect
DreamFect Gold
120
100
80
60
40
20
0
293T
J774
OZ Biosciences – DreamFect ™ Gold Transfection Reagent - Results – v1.2
CaCo2
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Higher Protein Expression Level with DreamFect Gold
DreamFect
DreamFect Gold
Relative Unit of Luciferase
500
450
400
350
300
250
200
150
100
50
0
Vero
HepG2
CV-1
Optimization of DNA /DreamFect Gold ratio
The general protocol is as simple as using 4 µL of DreamFect ™ Gold per 1 µg of DNA.
However, optimal conditions may vary depending on the nucleic acid, cell types, size of cell culture dishes and
presence or absence of serum. Therefore, the amounts and ratios of the individual components (DNA and
DreamFect Gold™) may have to be adjusted to achieve best results (see examples of results below).
Consequently, we suggest that you optimized this important parameter.
1.
2.
3.
4.
5.
The
The
The
The
The
ratio of DreamFect Gold / DNA
quantity of DNA
cell number
presence or absence of serum
incubation time
For instance, optimum DreamFect Gold / DNA ratio is 4/1 for A549, BEAS-2B, COS7, K562 and U87 in a 24-well
plate. Conversely, optimum ratio for HeLa, Raw, HEK293, CHO and BHK is 2/1 in a 24-well plate. Our team has
developed many cell type specific protocols with optimized transfection conditions. Please contact our technical
support service to obtain these protocols: [email protected]
Optimization of cell number and amount of DreamFect Gold
90
% Transfected Cells
80
70
5e+4 cells
10e+4 cells
20e+4 cells
60
50
40
30
20
10
0
1 µL
2 µL
4 µL
Volume of DreamFect Gold
Various numbers of A549 cells were transfected with 1 µg of pEGFP plasmid and 1 µL, 2µL or 4µL of DreamFect
Gold in a 24-well plate. The % of transfected cells were measured 24h post-transfection by cytofluorimetry.
OZ Biosciences – DreamFect ™ Gold Transfection Reagent - Results – v1.2
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Optimization of DNA amount
50
Quantity of β-galactosidase (ng)
45
COS7
A549
40
K562
35
30
25
20
15
10
5
0
0,125 µg
0,25 µg
0,5 µg
Quantity of DNA
CO7, A549 and K562 cells (15x103 cells / well) were transfected with various amounts of pLacZ plasmid and 0.5 µL, 1 µL and
2 µL of DreamFect ™ Gold reagent in a 96-well plate, respectively. β-Galactosidase expression was monitored 24 h after
transfection using OZ Biosciences’ ONPG β-galactosidase assay kit (catalog # G010001).
Bibliographic references
Please consult our list of references available on the website: www.ozbiosciences.com.
OZ Biosciences – DreamFect ™ Gold Transfection Reagent - Results – v1.2
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