Merck, CAS9RFPVP-1UG, Cas9-RFP Lenti Plasmid

General description
This product is a lentiviral plasmid that utilizes the EF1 alpha promoter to drive expression of Cas9 and RFP linked by a 2A peptide (EF1a-Cas9-2A-RFP) allowing for easy visualization of successful transfection or transduction. Use Sigma′s lentiviral Cas9 plasmid for generation of lentiviral particles and efficient production of stable cell lines expressing Cas9 for CRISPR based genome editing. Sigma′s Cas9-RFP lenti plasmid is one part of a two part CRISPR system with individual Cas9 and gRNA expression vectors. For easy fluorescence microscopy a MOI greater than 200 should be employed when packaged into lentiviral particles.

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Application
Functional Genomics/Target Validation
Creation of gene knockouts in multiple cell lines
Complete knockout of genes not amenable to RNAi
Manufacture of Cas9-RFP expressing Lentiviral Particles

Biochem/physiol Actions
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.
Features and Benefits
Highly specific
Highly active
Ready to use purified plasmid DNA

Legal Information
CRISPR Use License Agreement

Lentiviral, WPRE and Evrogen Fluorophore Licenses