Metabolic Incorporation of Azide Functionality into Cellular RNA

Chembiochem. 2016 Nov 17;17(22):2149-2152. doi: 10.1002/cbic.201600300. Epub 2016 Sep 30.

Abstract

Real-time tracking of RNA expression can provide insight into the mechanisms used to generate cellular diversity, as well as help determine the underlying causes of disease. Here we present the exploration of azide-modified nucleoside analogues and their ability to be metabolically incorporated into cellular RNA. We report robust incorporation of adenosine analogues bearing azide handles at both the 2'- and N6-positions; 5-methylazidouridine was not incorporated into cellular RNA. We further demonstrate selectivity of our adenosine analogues for transcription and polyadenylation. We predict that azidonucleosides will find widespread utility in examining RNA functions inside living cells, as well as in more complex systems such as tissues and living animals.

Keywords: RNA incorporation; azides; imaging; modified nucleosides; transcription.

MeSH terms

  • Adenosine / analogs & derivatives*
  • Adenosine / metabolism
  • Alkynes / chemistry
  • Azides / chemistry*
  • Catalysis
  • Copper / chemistry
  • Cycloaddition Reaction
  • Fluorescent Dyes / chemistry
  • HeLa Cells
  • Humans
  • Microscopy, Fluorescence
  • Nucleosides / chemistry*
  • RNA / chemistry
  • RNA / metabolism*
  • Ribonucleotide Reductases / antagonists & inhibitors

Substances

  • Alkynes
  • Azides
  • Fluorescent Dyes
  • Nucleosides
  • RNA
  • Copper
  • Ribonucleotide Reductases
  • Adenosine