Visualizing mRNA in live mammalian cells

Zejun Wang; Wenhan Liu; Chunhai Fan; Nan Chen

doi:10.1016/j.ymeth.2019.03.008

Visualizing mRNA in live mammalian cells

Methods. 2019 May 15:161:16-23. doi: 10.1016/j.ymeth.2019.03.008. Epub 2019 Mar 14.

Authors

Zejun Wang¹, Wenhan Liu², Chunhai Fan², Nan Chen³

Affiliations

¹ Department of Chemistry, Shanghai Normal University, Shanghai 200234, China; Division of Physical Biology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 201800, China.
² Division of Physical Biology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 201800, China.
³ Department of Chemistry, Shanghai Normal University, Shanghai 200234, China; Division of Physical Biology, Shanghai Institute of Applied Physics, Chinese Academy of Sciences, University of Chinese Academy of Sciences, Shanghai 201800, China. Electronic address: nchen@shnu.edu.cn.

PMID: 30878443
DOI: 10.1016/j.ymeth.2019.03.008

Abstract

mRNAs play essential role in gene expression and regulation. Direct visualization of specific intracellular mRNAs can provide key information about mRNA distribution and dynamics that accelerating study of their functionality. This highlights the necessity of characterizing mRNA in mammalian cells with good spatial and temporal resolution. Live cell imaging and tracking of mRNA remains a challenge due to their relatively short half-life and lack of versatile imaging probes. This review provides an overview of current methods of live-cell mRNA imaging, including exogenous hybridization probes and genetically encoded strategies. It then focuses on the development and application of aptamer initiated fluorescence complementation (AiFC) method for real-time imaging of mammalian mRNAs and includes a detailed protocol for observing β-actin mRNA in HeLa cells.

Publication types

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

MeSH terms

Animals
Base Sequence
Fluorescence Resonance Energy Transfer / methods*
Humans
Microscopy, Fluorescence / methods
RNA, Messenger / chemistry*
RNA, Messenger / genetics*

Substances

RNA, Messenger