The Dynamics of mRNA Turnover Revealed by Single-Molecule Imaging in Single Cells

Mol Cell. 2017 Nov 2;68(3):615-625.e9. doi: 10.1016/j.molcel.2017.09.030. Epub 2017 Oct 19.


RNA degradation plays a fundamental role in regulating gene expression. In order to characterize the spatiotemporal dynamics of RNA turnover in single cells, we developed a fluorescent biosensor based on dual-color, single-molecule RNA imaging that allows intact transcripts to be distinguished from stabilized degradation intermediates. Using this method, we measured mRNA decay in single cells and found that individual degradation events occur independently within the cytosol and are not enriched within processing bodies. We show that slicing of an mRNA targeted for endonucleolytic cleavage by the RNA-induced silencing complex can be observed in real time in living cells. This methodology provides a framework for investigating the entire life history of individual mRNAs from birth to death in single cells.

Keywords: P-bodies; RNAi; Xrn1; fluorescence microscopy; live-cell imaging; mRNA decay; single-molecule.

Publication types

  • Video-Audio Media

MeSH terms

  • Argonaute Proteins / genetics
  • Argonaute Proteins / metabolism
  • Gene Expression Regulation
  • Genes, Reporter
  • HeLa Cells
  • Humans
  • Kinetics
  • Microscopy, Fluorescence*
  • Microscopy, Video
  • Models, Genetic
  • RNA Stability*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*
  • Single Molecule Imaging / methods*
  • Transfection


  • AGO2 protein, human
  • Argonaute Proteins
  • RNA, Messenger