47 Life Technologies
™
| Genome editing
0
5
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35
40
Lipofectamine
®
MessengerMAX
™
reagent
Competitor A Competitor B Lipofectamine
®
3000 Cells only
Cleavage
efficiency
(%)
Cas9 mRNA + IVT gRNA
All-in-one CRISPR plasmid
GeneArt
®
CRISPR
Nuclease mRNA
In vitro transcribe gRNAs
Use MEGAshortscipt
™
T7 Transcription Kit
for gRNA synthesis
IVT gRNAs
TRANSFECT
with Lipofectamine
®
MessengerMAX
™
reagent
Up to 10x higher cleavage efficiency with mRNA CRISPRs
Lipofectamine
®
MessengerMAX
™
reagent helps increase the likelihood of cleavage
and recombination with GeneArt
®
CRISPR Nuclease mRNA through highly efficient
transfection, maximizing the efficiency of genetic modifications and simplifying the
downstream processes (Figure 12).
Figure 12 . Cleavage efficiency of various GeneArt
®
CRISPR formats targeting the HPRT locus in Gibco
®
iPS cells
in a 12-well format. Lipofectamine
®
3000 reagent was used to deliver CRISPR nuclease all-in-one plasmid DNA;
Lipofectamine
®
MessengerMAX
™
and two leading mRNA delivery reagents were used to deliver an all-RNA CRISPR
format (Cas9 mRNA + IVT gRNA). Cleavage efficiency was determined using the GeneArt
®
Genomic Cleavage Detection
Kit 72 hours posttransfection.
