General description

This product is an expression plasmid that utilizes the CMV promoter for strong transient expression of Dead Cas9 (CMV-dCas9). The dCas9 expression plasmid is one part of a two part CRISPR system with individual dCas9 and gRNA expression vectors.

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Application

Functional Genomics/Target Validation

Proxy CRISPR
Suitable for genomic DNA detection

Features and Benefits

Highly specific
Sequence verified
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 rendered inactive (dCas9) with mutations to the two protein domains, RuvC and HnH (D10A and H840A respectively), which are responsible for nuclease activity. The nuclease deficient protein can then be programmed with a gRNA and directed to bind at a desired sequence of DNA. Variants of programmable endonucleases are often unable to cleave a large number of the targets that are efficiently cleaved by canonical SpCas9 in human cells. Cotargeting dead Cas9 may alter local chromatin structures and render otherwise inaccessible target sites now accessible and cleavable by alternative nucleases such as type II-B FnCas9, type II-C CjCas9 and NcCas9 and type V FnCpf1.