CleanCap M6 Cas9 mRNA (N1MePsU), 1 mg

Cas9 mRNA expresses a version of theStreptococcus pyogenesSF370 Cas9 protein (CRISPR Associated Protein 9) . Cas9 functions as part of the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats) genome editing system. In the CRISPR system, an RNA guide sequence targets the site of interest and the Cas9 protein is employed to perform double-stranded DNA cleavage. Cas9 mRNA encodes the Cas9 protein with an N and C terminal nuclear localization signal (NLS) . The incorporation of two NLS signals within the mRNA increases the frequency of delivery to the nucleus, thus increasing the rate of DNA cleavage. Additionally, a C terminal HA epitope tag aids in detection, isolation, and purification of the Cas9 protein. This mRNA is capped usingCleanCap® Reagent M6, TriLink's patented co-transcriptional capping technology, resulting in the naturally occurring Cap-1 structure with >95% capping efficiency. It is polyadenylated, modified withN1-methylpseudouridine, and optimized for mammalian systems. It mimics a fully processed mature mRNA. CleanCap Reagent M6, otherwise known as CleanCap m6AG 3' OMe, produces a base-modified Cap-1 mRNA, which shows superior in vivo activity compared to Cap-0 mRNA produced by legacy capping methods such as mCap or anti-reverse cap analog (ARCA) . CleanCap M6 may further increase protein expression relative to previous generations of cap analogs, such as CleanCap AG or CleanCap AG (3' OMe), or mRNAs produced by enzymatic capping strategies. N1-methylpseudouridine is a modified uridine that can reduce immunogenic response and enhance translational efficiency of mRNAs. These properties can result in safer mRNA and increased protein expression.