The ScriptCap™ 2’-O-Methyltransferase, Vaccinia Virus Cap 1 methyltransferase (VMT), provides a convenient method to prepare fully cap 1- capped RNA in vitro from any source of cap 0-capped RNA.1 Caps play a crucial role in maintaining the stability and translational efficiency of mRNA in vivo. Cap 1 methylation serves, at least in part, to increase the translation efficiency of the mRNA.1,2 VMT activity is dependent on the presence of a 5'-cap 0 cap structure (m7G[5']ppp[5']-NpN...). Once the RNA is capped, VMT catalyzes the transfer of a methyl group from the donor molecule S-adenosyl-methionine (SAM) to the 2'-O position of the penultimate nucleotide forming a cap 1 cap structure (m7Gppp[m2’-O]NpNpN...).3-5 While cap 0-capped RNA can be produced by use
of a dinucleotide cap analog in an in vitro transcription system, cap 1-capped RNA cannot be produced.6 The VMT substrate can be cap 0-capped RNA generated from the ScriptCap™ m7G Capping System (produced with a capping enzyme) 7 or cap analog-based transcription kits such as the AmpliCap-Max™ High Yield Message Maker Kits. The ScriptCap 2'-O-Methyltransferase can be
used simultaneously with the ScriptCap m7G Capping System or sequentially following cap analogbased transcription reactions. ScriptCap m7G Capping System / 2'-O-Methyltransferase reactions
can added directly to Poly(A) Polymerase reactions for convenient 3'-end poly(A)-tailing of the capped RNA.