NLS-Cas9-D10A Nickase Self-Amplifying mRNA

This self-amplifying mRNA encodes four nonstructural proteins (nsP1 to nsP4) derived from a positive-stranded alphavirus venezuelan equine encephalitis virus (VEEV), and NLS-Cas9-D10A Nickase.

NLS-Cas9-D10A Nickase is a Streptococcus pyogenes SF370 Cas9 protein D10A mutant with nuclear localization signal (NLS) at C terminus. Unlike the wild-type CAS9, which cleaves the double strand DNA generating two break sites based on its two active domains, the NLS-Cas9-D10A Nickase has only one active domain, thus it can only cut one single strand DNA that is complementary to the guide-RNA, producing one single strand break. The mutant mRNA favors homology-directed repair and decreases the occurrence of non-homologous end joining.

• 5' Capped with vaccinia capping enzyme system and 2'-O-methyltransferase to yield cap-1 structure
• Pseudo-UTP
• 5mCTP
• PolyA: 180 nt