Cas9-GFP Lenti Plasmid CAS9GFPVP-1UG

This product is a lentiviral plasmid that utilizes the EF1 alpha promoter to drive expression of Cas9 and GFP linked by a 2A peptide (EF1a-Cas9-2A-GFP) allowing for easy visualization of successful transfection or transduction. Use Sigmas lentiviral Cas9 plasmid for generation of lentiviral particles and efficient production of stable cell lines expressing Cas9 for CRISPR based genome editing. Sigmas Cas9-GFP lenti plasmid is one part of a two part CRISPR system with individual Cas9 and gRNA expression vectors.
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Functional Genomics/Target Validation

• Creation of gene knockouts in multiple cell lines
• Complete knockout of genes not amenable to RNAi
• Manufacture of Cas9-GFP expressing Lentiviral Particles

Features and Benefits

• Highly specific
• Highly active
• Ready to use purified plasmid DNA

Principle
CRISPR/Cas systems are employed by bacteria and archaea as a defense against invading viruses and plasmids. Recently, the type II CRISPR/Cas system from the bacterium Streptococcus pyogenes has been engineered to function in eukaryotic systems using two molecular components: a single Cas9 protein and a non-coding guide RNA (gRNA). The Cas9 endonuclease can be programmed with a single gRNA, directing a DNA double-strand break (DSB) at a desired genomic location. Similar to DSBs induced by zinc finger nucleases (ZFNs), the cell then activates endogenous DNA repair processes, either non-homologous end joining (NHEJ) or homology-directed repair (HDR), to heal the targeted DSB.
CRISPR Use License Agreement,
Lentiviral, WPRE and Evrogen Fluorophore Licenses,

recombinant	expressed in E. coli
packaging	vial of 50 µL
concentration	20 ng/µL in TE buffer; DNA (1µg of plasmid DNA)
reporter gene	GFP
selection	ampicillin
shipped in	dry ice
storage temp.	20°C

RIDADR	NONH for all modes of transport
WGK Germany	WGK 2
Flash Point F	Not applicable
Flash Point C	Not applicable