Synthetic mRNA capping

• Fabian Muttach,
• Nils Muthmann and
• Andrea Rentmeister

Beilstein J. Org. Chem. 2017, 13, 2819–2832, doi:10.3762/bjoc.13.274

• ; enzymatic capping; methyltransferase; RNA; Introduction The 5′-cap is a hallmark of eukaryotic mRNA and involved in numerous interactions required for cellular functions. Chemically, the 5′-cap consists of an inverted 7-methylguanosine connected to the rest of the eukaryotic mRNA via a 5′–5′ triphosphate

• -transcriptional capping In post-transcriptional capping, the RNA from IVT is subjected to a dedicated enzymatic capping reaction. The enzymes used in vitro originate from capping apparatuses of different eukaryotic organisms or DNA viruses and can be produced recombinantly in E. coli [28][29]. Enzymatic formation

• ]. This was achieved by combining solid-phase synthesis and enzymatic modification. Specifically, 5′-diphosphorylated RNAs (up to 170 nt long) were chemically synthesized, cleaved from the solid support, deprotected and purified. This was followed by enzymatic capping, 2′-O-methylation and polyadenylation