eSpCas9-Blasticidin Lenti Plasmid

Description

• General description: 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 Sigma′s lentiviral eSpCas9 plasmid for generation of lentiviral particles and efficient production of stable cell lines expressing eSpCas9 for CRISPR based genome editing. Sigma′s lenti-eSpCas9 plasmid is one part of a two part CRISPR system with individual eSpCas9 and gRNA expression vectors.To order gRNA in any format click here
• Application: Functional Genomics/Target ValidationCreation of gene knockouts in multiple cell linesComplete knockout of genes not amenable to RNAiManufacture of eSpCas9 expressing Lentiviral Particles
• Features and Benefits: Enhanced specificity compared to wild type Cas9Highly ActiveReady 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.[1]
• Legal Information: CRISPR Use License AgreementLentiviral, WPRE and Evrogen Fluorophore Licenses