Description
In eukaryotes, post-transcriptionally modified mRNA forms a special cap structure (m7Gppp5'N) at the 5' end, which is closely related to mRNA stability, transport, and translation. As a simple and effective method, enzymatic capping can significantly improve the stability and translation ability of in vitro transcribed RNA. Vaccinia capping enzyme is composed of two subunits (D1, D12) with RNA triphosphatase activity, guanylyltransferase activity, and guanine methyltransferase activity, which are required for adding Cap0 structure. In the presence of appropriate concentrations of S-Adenosylmethionine (SAM), guanosine triphosphate (GTP), etc., vaccinia capping enzyme can add cap structure (m7Gppp, Cap0) to the 5' end of RNA. Based on the enzymatic reaction, the capping efficiency can be close to 100%, and the cap structure is oriented correctly