Firefly luciferase saRNA

Firefly luciferase, originally extracted from firefly Photinus pyralis, can catalyze the oxidation of substrate D-luciferin to emit light at a wavelength of about 560nm. Firefly luciferase is a commonly used reporter gene, which is often used in the study of gene regulation and function.

Self-Amplifying RNA(saRNA) usually has the advantages of high protein expression, long duration, and low dose use due to its ability to self-replicate within the cell. Firefly luciferase saRNA is based on the replication mechanism of Venezuelan equine encephalitis virus (VEEV)，providing a concentration of approximately 1 mg/mL with Cap1 structure. There are currently two ways to cap mRNA: One is co-transcription method, by adding Cap analogues into the transcription process. The other is enzymatic Capping. After transcription, Cap0 capping is performed by Vaccinia virus Capping Enzyme (VCE), GTP and S-adenosylmethionine (SAM). The Cap0 is then generated into the Cap1 through 2´-O-Methyltransferase and SAM. Cap1 Capping can also be performed by adding VCE, 2´-O-Methyltransferase, GTP and SAM in a one-step process. Cap 1 structure is more ideal for mammalian systems and possess higher transcription efficiency than Cap 0 structure. The addition of poly (A) tail increases the stability and lifetime of the mRNA in vitro and in vivo. Poly (A) tail also plays an important role in enhancing the efficiency of translation initiation. Firefly Luciferase saRNA can stable expression of firefly luciferase for a long time, suitable for transfection, cell viability and in vivo imaging experiments.