Oligonucleotide-Addressed Covalent 3'-Terminal Derivatization of Small RNA Strands for Enrichment and Visualization

Angew Chem Int Ed Engl. 2017 Jun 1;56(23):6507-6510. doi: 10.1002/anie.201701448. Epub 2017 May 4.

Abstract

The HEN1 RNA 2'-O-methyltransferase plays important roles in the biogenesis of small non-coding RNAs in plants and proved a valuable tool for selective transfer of functional groups from cofactor analogues onto miRNA and siRNA duplexes in vitro. Herein, we demonstrate the versatile HEN1-mediated methylation and alkylation of small RNA strands in heteroduplexes with a range of complementary synthetic DNA oligonucleotides carrying user-defined moieties such as internal or 3'-terminal extensions or chemical reporter groups. The observed DNA-guided covalent functionalization of RNA broadens our understanding of the substrate specificity of HEN1 and paves the way for the development of novel chemo-enzymatic tools with potential applications in miRNomics, synthetic biology, and nanomedicine.

Keywords: HEN1; methyltransferases; microRNA; siRNA; synthetic AdoMet analogues.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alkylation
  • Methylation
  • Methyltransferases / metabolism
  • MicroRNAs / chemistry*
  • Nucleic Acid Heteroduplexes / chemistry
  • Oligonucleotides / chemistry*
  • RNA, Small Interfering / chemistry*
  • RNA, Small Untranslated / chemistry*
  • Substrate Specificity

Substances

  • MicroRNAs
  • Nucleic Acid Heteroduplexes
  • Oligonucleotides
  • RNA, Small Interfering
  • RNA, Small Untranslated
  • Methyltransferases
  • RNA 2'-O-methyltransferase