Selective covalent labeling of miRNA and siRNA duplexes using HEN1 methyltransferase

J Am Chem Soc. 2014 Oct 1;136(39):13550-3. doi: 10.1021/ja507390s. Epub 2014 Sep 19.

Abstract

MicroRNAs regulate gene expression in numerous biological pathways and are typically methylated at their 3'-termini in plants but not in animals. Here we show that the HEN1 RNA 2'-O-methyltransferase from Arabidopsis thaliana catalyzes the transfer of extended propargylic moieties from synthetic AdoMet cofactor analogs to duplex miRNAs or siRNAs. The presented approach permits selective and efficient covalent labeling of small RNA duplexes with a variety of functional or reporter groups for their enrichment and analysis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / enzymology*
  • Arabidopsis Proteins / metabolism*
  • Biocatalysis
  • Methyltransferases / metabolism*
  • MicroRNAs / chemistry
  • MicroRNAs / metabolism*
  • Molecular Structure
  • RNA, Small Interfering / chemistry
  • RNA, Small Interfering / metabolism*
  • Staining and Labeling / methods*

Substances

  • Arabidopsis Proteins
  • HEN1 protein, Arabidopsis
  • MicroRNAs
  • RNA, Small Interfering
  • Methyltransferases
  • RNA 2'-O-methyltransferase