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A transgenic mouse for in vivo detection of endogenous labeled mRNA.

Live-cell single mRNA imaging is a powerful tool but has been restricted in higher eukaryotes to artificial cell lines and reporter genes. We describe an approach that enables live-cell imaging of single endogenous labeled mRNA molecules transcribed in primary mammalian cells and tissue. We generated a knock-in mouse line with an MS2 binding site (MBS) cassette targeted to the 3' untranslated region of the essential β-actin gene. As β-actin-MBS was ubiquitously expressed, we could uniquely address endogenous mRNA regulation in any tissue or cell type. We simultaneously followed transcription from the β-actin alleles in real time and observed transcriptional bursting in response to serum stimulation with precise temporal resolution. We tracked single endogenous labeled mRNA particles being transported in primary hippocampal neurons. The MBS cassette also enabled high-sensitivity fluorescence in situ hybridization (FISH), allowing detection and localization of single β-actin mRNA molecules in various mouse tissues.

Pubmed ID: 21240280


  • Lionnet T
  • Czaplinski K
  • Darzacq X
  • Shav-Tal Y
  • Wells AL
  • Chao JA
  • Park HY
  • de Turris V
  • Lopez-Jones M
  • Singer RH


Nature methods

Publication Data

February 31, 2011

Associated Grants

  • Agency: NIBIB NIH HHS, Id: EB2060
  • Agency: NIGMS NIH HHS, Id: F32-GM087122
  • Agency: NIGMS NIH HHS, Id: GM84364
  • Agency: NIGMS NIH HHS, Id: GM86217
  • Agency: NIBIB NIH HHS, Id: R01 EB002060
  • Agency: NIBIB NIH HHS, Id: R01 EB002060-25
  • Agency: NIGMS NIH HHS, Id: R01 GM084364
  • Agency: NIGMS NIH HHS, Id: R01 GM084364-18
  • Agency: NIGMS NIH HHS, Id: R01 GM086217
  • Agency: NIGMS NIH HHS, Id: R01 GM086217-04

Mesh Terms

  • Animals
  • Cell Survival
  • Cells, Cultured
  • In Situ Hybridization, Fluorescence
  • Mice
  • Mice, Transgenic
  • RNA, Messenger
  • Transcription, Genetic