Current covalent modification methods for detecting RNA in fixed and living cells

Methods. 2016 Apr 1:98:18-25. doi: 10.1016/j.ymeth.2015.11.016. Epub 2015 Nov 23.

Abstract

Labeling RNAs is of particular interest for elucidating localization, transport, and regulation of specific transcripts, ideally in living cells. Numerous methods have been developed ranging from hybridizing probes to genetically encoded reporters and chemo-enzymatic approaches. This review focuses on covalent labeling approaches that rely on the introduction of a small reactive group into the nascent or completed transcript followed by bioorthogonal click chemistry. State of the approaches for labeling RNA in fixed and living cells will be presented and emerging strategies with great potential for application in the complex cellular environment will be discussed.

Keywords: Chemo-enzymatic; Click chemistry; Labeling; RNA visualization.

Publication types

  • Research Support, Non-U.S. Gov't
  • Review

MeSH terms

  • Base Pairing
  • Cell Line
  • Cellular Microenvironment
  • Click Chemistry / methods*
  • Fluorescent Dyes / chemistry
  • Humans
  • MicroRNAs / chemistry*
  • MicroRNAs / genetics
  • MicroRNAs / metabolism
  • Molecular Probes / chemistry*
  • Oligonucleotides / chemical synthesis
  • Oligonucleotides / chemistry
  • RNA, Messenger / chemistry*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Single Molecule Imaging / instrumentation
  • Single Molecule Imaging / methods*
  • Solid-Phase Synthesis Techniques / methods
  • Staining and Labeling / methods*
  • Tissue Fixation / methods
  • Transcription, Genetic

Substances

  • Fluorescent Dyes
  • MicroRNAs
  • Molecular Probes
  • Oligonucleotides
  • RNA, Messenger