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Mol Cell. 2018 Aug 2;71(3):468-480. doi: 10.1016/j.molcel.2018.07.022.

A Growing Toolbox to Image Gene Expression in Single Cells: Sensitive Approaches for Demanding Challenges.

Author information

1
Institut de Génétique Moléculaire de Montpellier, Université de Montpellier, CNRS, Montpellier, France; Equipe labélisée Ligue Nationale Contre le Cancer, Montpellier, France.
2
Institut de Génétique Moléculaire de Montpellier, Université de Montpellier, CNRS, Montpellier, France.
3
Unité Imagerie et Modélisation, Institut Pasteur and CNRS UMR 3691, 28 rue du Docteur Roux, 75015 Paris, France.
4
Institut de Génétique Moléculaire de Montpellier, Université de Montpellier, CNRS, Montpellier, France; Equipe labélisée Ligue Nationale Contre le Cancer, Montpellier, France; Department of Anatomy and Structural Biology, Albert Einstein College of Medicine, Bronx, NY, USA. Electronic address: edouard.bertrand@igmm.cnrs.fr.

Abstract

The spatiotemporal regulation of gene expression is key to many biological processes. Recent imaging approaches opened exciting perspectives for understanding the intricate mechanisms regulating RNA metabolism, from synthesis to decay. Imaging techniques allow their observation at high spatial and temporal resolution, while keeping cellular morphology and micro-environment intact. Here, we focus on approaches for imaging single RNA molecules in cells, tissues, and embryos. In fixed cells, the rapid development of smFISH multiplexing opens the way to large-scale single-molecule studies, while in live cells, gene expression can be observed in real time in its native context. We highlight the strengths and limitations of these methods, as well as future challenges. We present how they advanced our understanding of gene expression heterogeneity and bursting, as well as the spatiotemporal aspects of splicing, translation, and RNA decay. These insights yield a dynamic and stochastic view of gene expression in single cells.

KEYWORDS:

MS2; RNA imaging; RNA processing; gene expression; noise; single molecule; smFISH; splicing; transcription; translation

PMID:
30075145
DOI:
10.1016/j.molcel.2018.07.022
[Indexed for MEDLINE]
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