Synthesis of cap-2 mrna

Abstract

Described herein is method of synthesizing a Cap-2 mRNA molecule comprises co-transcriptionally installing a priming trinucleotide ppAmAmG to a 5′-terminus of an mRNA molecule during an in vitro transcription reaction, wherein Am denotes 2′-O-methyladenosine, and adding m7G to the ppAmAmG using a vaccinia capping enzyme to yield a Cap-2 mRNA molecule comprising m7GpppAmAmG at the 5′-terminus. A trinucleotide compound having the structure ppAmAmG, wherein pp denotes a diphosphate moiety at a 5′-terminus, and wherein Am denotes 2′-O-methyladenosine, or a pharmaceutically acceptable salt thereof, is also provided. A method of inducing an immune response in a subject comprises administering to the subject a Cap-2 mRNA molecule comprising m7GpppAmAmG at a 5′-terminus, wherein the Cap-2 mRNA molecule encodes an antigen.

Dependencies are inferred from the claim text.