EZ Cap™ Cas9 mRNA (5-moUTP)

The Cas9/guide RNA (Cas9/gRNA) system is commonly used for genome editing. Upon delivery of these components to the nucleus of a cell, the RNA guide sequence targets the site of interest and the Cas9 protein is employed to perform the DNA cleavage.

EZ Cap™ Cas9 mRNA (5-moUTP) is provided at a concentration of ~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 5-moUTP and poly(A) tail suppress RNA-mediated innate immune activation and increase 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.