® GenomONE series HVJ Envelope transfection and cell fusion kits ISHIHARA SANGYO KAISHA, LTD. https://www.iskweb.co.jp/eng/products/hvj-e/ Genome Editing Membrane fusion-mediated intracellular delivery of Cas9 protein enables efficient genome editing even in mouse primary T cells. Applicable to CRISPR-Cas9-mediated knock-in and knock-out Higher genome editing efficiency than that of the competitors Transfection Highly efficient transfection of plasmid DNA, siRNA/miRNA, and protein Applicable to immune cells (e.g., Jurkat, THP-1, U-937, RAW 264.7, and mouse primary T cells) Applicable to in vivo transfection Cell fusion Alternative method to PEG and electrofusion for cell fusion Simple operability, low cytotoxicity and high fusion activity Applicable to hybridoma preparation and nuclear transfer
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HVJ Envelope transfection and cell fusion kits GenomONEseries · Transfection Highly efficient transfection of plasmid DNA, siRNA/miRNA, and protein Applicable to immune cells (e.g.,
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®
GenomONE series
HVJ Envelope transfection and cell fusion kits
ISHIHARA SANGYO KAISHA, LTD.
https://www.iskweb.co.jp/eng/products/hvj-e/
Genome EditingMembrane fusion-mediated intracellular delivery of Cas9 protein enables
efficient genome editing even in mouse primary T cells.
・Applicable to CRISPR-Cas9-mediated knock-in and knock-out
・Higher genome editing efficiency than that of the competitors
TransfectionHighly efficient transfection of plasmid DNA, siRNA/miRNA, and protein
・Applicable to immune cells (e.g., Jurkat, THP-1, U-937, RAW 264.7, and mouse primary T cells)
・Applicable to in vivo transfection
Cell fusionAlternative method to PEG and electrofusion for cell fusion
・Simple operability, low cytotoxicity and high fusion activity
・Applicable to hybridoma preparation and nuclear transfer
GenomONE series
GenomONE-NeoPlasmid DNA, siRNA/miRNA, protein transfection kit (in vivo)
High knockdown effects were obtained using GenomONE-Si, whereas no adequate effect could be obtained using other transfection
reagents; thus, the superiority of GenomONE-Si was demonstrated.
CDC2 siRNA
a; Negative control miRNA (1000pM)b; Cyclophilin B miRNA (1000pM)
c; Cyclophilin B miRNA (400pM)d; Cyclophilin B miRNA (100pM)
e; Cyclophilin B miRNA (25pM)
f ; Cyclophilin B miRNA (6.3pM)
β-actin
Cyclophilin B
a b c d e f
Western blotting
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1.2
Control(PBS)
400pM 400pM 100pM 25pM 6.3pM
N.C.miRNA
Cyclophilin B miRNA
Ce
ll Via
bility
(%)
Re
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Qu
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(no
rma
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d 1
8S
rR
NA
)
Relative Quantity Cell Viability(%)
75%
56%
Product X
Product R
GenomONE-Si
Re
lati
ve
Qu
an
tity
(no
rma
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d 1
8S
rR
NA
)
Re
lati
ve
Qu
an
tity
(no
rma
lize
d 1
8S
rR
NA
)
GenomONE-SiGenomONE-Si
®
・Optimal for transfection of synthetic oligo-type siRNA/miRNA
・Applicable to difficult-to-transfect immune cells
・Optimal for rapid screening of a large number of test samples (high-throughput screening)
KALA peptides can destabilize the endosomal membrane and
enhance the transfection efficiency of non-viral gene delivery
vectors.
KALA peptide: WEAKLAKALAKALAKHLAKALAKALKACEA
GenomONE - GX・A novel gene delivery vector composed of a lipid-like substance and HVJ-E
・The GenomONE-GX Enhancer suppresses innate immune responses by plasmid DNA and
improves transgene expression.
・Transgene expression is enhanced by GenomONE-GX using a combination of KALA peptides.
■■■■ TurboGFP expression
Various cell lines were transfected with a reporter plasmid encoding TurboGFP under a CAG promoter using either GenomONE-GX(with or without the KALA peptide) or Product L (a competitor), following the manufacturer’s instructions.
※The KALA peptide is not included in the GenomONE-GX kit and hence the KALA peptide solution needs to be prepared in advance.
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GenomONE-GX
Product L
(competitor)
Jurkat K-562 RAW 264.7 HEK-293 RIN-5F SH-SY5Y
GenomONE-GX
+KALA peptide
■■■■ Mechanism of Enhancer
The GenomONE-GX Enhancer inhibits innate immune signaling
in response to the recognition of exogenous DNA. Hence, it
can be used to increase transgene expression, which is
intrinsically suppressed by innate immune signaling.
35 copies of PCAG-loxP-neopA-loxP-LacZ were tandemly inserted into the genome.
Cre recombinase
alone
HVJ-E
+Cre recombinase
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GenomONE - CF・A cell fusion reagent with low cytotoxicity and
high fusion activity・Applicable to hybridoma preparation and
nuclear transfer・An alternative method to PEG and electrofusion for cell fusion
■■■■ Comparison with PEG method in the fusion of different types of cell
■■■■ Comparison with PEG method in hybridoma preparation
Rat MSC cells (rat bone marrow-derived mesenchymal stem cells) labeled with red fluorescence were combined with rat primary
cardiac myocytes labeled with green fluorescence in Cell Fusion Buffer. As a result, fused cells (yellow) were formed (GenomONE-CFsuspension method). Fused cells adhering to the plate were also observed after 1-2 days of culturing. In the PEG-treated group, high
cytotoxicity appeared immediately after cell fusion, reducing the number of fused cells obtained.
Normal BALB/c mouse splenocytes not sensitized with antigen were fused to X63-Ag8.653 myeloma cells using GenomONE -CF or
PEG1500. Beginning the following day, half of the culture medium (10%FBS/RPMI1640) was replaced with HAT medium at five
points of time(Days 1, 2, 3, 5, and 8), and the growth of colonies in each well was assessed on Days 10 - 11 to determine the
hybridoma-positive rate(an indicator of efficiency of fusion). On Day 12, mouse antibody level (IgG + IgA + IgM) in the supernatant
was measured by ELISA, to calculate the antibody production-positive rate. The effect of adding a commercially available hybridoma
supplement to the medium after fusion was also assessed (supplement was also added to the HAT medium).
Use of GenomONE -CF resulted in more efficient formation of antibody-producing hybridoma than that of PEG. The efficiency of cell
fusion mediated by GenomONE -CF was increased by the addition of hybridoma supplement to the medium used for incubation after
cell fusion.
Immediately after mixing
GenomONE-CF
PEG
®
After one day of culturing After 2 days of culturing
Hybridoma supplement
Hybridoma-positive rate
Antibody production-
positive rate
GenomONE -CF
-
--
-38/96 (40%) 9/96 (9%)
+
++
+96/96 (100%) 96/96 (100%)
PEG1500-
--
-3/96 (3%) 1/96 (1%)
+
++
+36/96 (38%) 9/96 (9%)
GenomONE series
Product selection guide
Product information
Visit our website for more detailed information regarding applications,
These products are sold for research purpose only. It may not be used for treatment or other clinical purposes or for intra- and extracorporeal diagnosis in humans or animals.