Tracking Distinct RNA Populations Using Efficient and Reversible Covalent Chemistry

Mol Cell. 2015 Sep 3;59(5):858-66. doi: 10.1016/j.molcel.2015.07.023.


We describe a chemical method to label and purify 4-thiouridine (s(4)U)-containing RNA. We demonstrate that methanethiosulfonate (MTS) reagents form disulfide bonds with s(4)U more efficiently than the commonly used HPDP-biotin, leading to higher yields and less biased enrichment. This increase in efficiency allowed us to use s(4)U labeling to study global microRNA (miRNA) turnover in proliferating cultured human cells without perturbing global miRNA levels or the miRNA processing machinery. This improved chemistry will enhance methods that depend on tracking different populations of RNA, such as 4-thiouridine tagging to study tissue-specific transcription and dynamic transcriptome analysis (DTA) to study RNA turnover.

Publication types

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

MeSH terms

  • Biotin / analogs & derivatives
  • Cell Proliferation
  • Disulfides
  • Gene Expression Profiling / methods
  • HEK293 Cells
  • Humans
  • Indicators and Reagents
  • Mesylates
  • MicroRNAs / chemistry*
  • MicroRNAs / genetics
  • MicroRNAs / metabolism
  • Organic Chemistry Phenomena
  • RNA Processing, Post-Transcriptional
  • Thiouridine / chemistry


  • Disulfides
  • Indicators and Reagents
  • Mesylates
  • MicroRNAs
  • N-(6-(biotinamido)hexyl)-3'-(2'-pyridyldithio)propionamide
  • Thiouridine
  • methanethiosulfonate
  • Biotin