SpCas9 mRNA (ARCA, 5mCTP, ψUTP)

SpCas9 mRNA expresses an optimized version of Streptococcus pyogenes protein. It can be used with a purified guideRNA sequence for genomic DNA target recognition and cleavage in the CRISPR-Cas genome editing system. Type II CRISPR-Cas system from the bacterium S. pyogenes has been adapted for genome engineering in eukaryotic systems, which requires two main components: the endoculease Cas9 and non-coding guide RNA (gRNA). The Cas9 protein can be programmed with gRNA, targeting a site-specific DNA double-strand break (DSB). Our SpCas9 mRNA is provided at a concentration of 1mg/ml, it is ARCA capped, polyA-tailed and incorporated with modified nucleotides. The capping of ARCA (anti-reverse cap analog) ensures high translation efficacy, while the addition of 5mCTP/ψUTP and poly(A) tail reduces host cell immune response and enhance mRNA stability. Moreover, Cas9 mRNA expresses the Cas9 protein with two nuclear localization signal (NLS) and C terminal HA epitope tag that enhance the rate of DNA cleavage and facilitate downstream purification.