NLS-FLP Recombinase Self-Amplifying mRNA, SA-NLS-FLP

FLP recombinase catalyzes the site-specific recombination of DNA between FRT sites. It has been widely used for generating conditional knockout mice, mediated by FLP/FRT system. Codon optimized FLP is a human/mouse codon-optimized thermo-stable mutant form of FLP, with enhanced recombinase activity by 4-10 fold higher than FLP at 37°C.

The self-amplifying NLS-Flp mRNA (SA-NLS-Flp) encodes four nonstructural proteins (NSP1 to NSP4) derived from a positive-stranded alphavirus venezuelan equine encephalitis virus (VEEV), in addition to GOI NLS-Flp. Upon entering the cytoplasm, the mRNA is translated to produce replicase complex, which amplifies the original NLS-Flp mRNA template to produce numerous copies of itself (mRNA replication), thus driving robust NLS-Flp protein expression.

SA-NLS-Flp mRNA induces much higher ex vivo recombination efficiency in cells than NLS-Flp mRNA with lower dose of delivery.

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

• Agarose gel electrophoresis
• Activity assay: HEK293 cells transfected with FRT reporter plasmid