Combining Chemical Synthesis and Enzymatic Methylation to Access Short RNAs with Various 5' Caps. Academic Article uri icon

Overview

abstract

  • Eukaryotic RNAs are heavily processed, including co- and post-transcriptional formation of various 5' caps. In small nuclear RNAs (snRNAs) or small nucleolar RNAs (snoRNAs), the canonical 7m G cap is hypermethylated at the N2 -position, whereas in higher eukaryotes and viruses 2'-O-methylation of the first transcribed nucleotide yields the cap1 structure. The function and potential dynamics of several RNA cap modifications have not been fully elucidated, which necessitates preparative access to these caps. However, the introduction of these modifications during chemical solid-phase synthesis is challenging and enzymatic production of defined short and uniform RNAs also faces difficulties. In this work, the chemical synthesis of RNA is combined with site-specific enzymatic methylation by using the methyltransferases human trimethylguanosine synthase 1 (hTgs1), trimethylguanosine synthase from Giardia lamblia (GlaTgs2), and cap methyltransferase 1 (CMTR1). It is shown that RNAs with di-and trimethylated caps, as well as RNAs with caps methylated at the 2'-O-position of the first transcribed nucleotide, can be conveniently prepared. These highly modified RNAs, with a defined and uniform sequence, are hard to access by in vitro transcription or chemical synthesis alone.

publication date

  • May 27, 2019

Research

keywords

  • Methyltransferases
  • RNA Cap Analogs

Identity

PubMed Central ID

  • PMC6755138

Scopus Document Identifier

  • 85069966070

Digital Object Identifier (DOI)

  • 10.1038/s41589-019-0231-8

PubMed ID

  • 30768827

Additional Document Info

volume

  • 20

issue

  • 13