Description |
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.
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