eSpCas9-Blasticidin Lenti Plasmid ESPCAS9BST-1EA

This product is a lentiviral plasmid that utilizes the EF1 alpha promoter to drive expression of eSpCas9 and a blasticidin resistance cassette linked by a 2A peptide (EF1a-eSpCas9-2A-Blasticidin) allowing for easy selection following successful transfection or transduction. Use Sigmas lentiviral eSpCas9 plasmid for generation of lentiviral particles and efficient production of stable cell lines expressing eSpCas9 for CRISPR based genome editing. Sigmas lenti-eSpCas9 plasmid is one part of a two part CRISPR system with individual eSpCas9 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 eSpCas9 expressing Lentiviral Particles

Features and Benefits

• Enhanced specificity compared to wild type Cas9
• 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. Newly engineered eSpCas9 enables the efficient targeted gene editing of established CRISPR systems with the benefit of reduced off-target effects. Point mutations in the chromosome-binding motif of SpCas9, as described by Slaymaker, et al., provide higher on-target fidelity without loss of cleavage efficiency.
CRISPR Use License Agreement,
Lentiviral, WPRE and Evrogen Fluorophore Licenses,

Quality Level	200,
recombinant	expressed in E. coli
packaging	vial of 50 µL
concentration	20 ng/µL in TE buffer; DNA (1µg of Plasmid DNA)
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