Live-cell imaging of endogenous mRNAs with a small molecule

Shin Ichi Sato; Mizuki Watanabe; Yousuke Katsuda; Asako Murata; Dan Ohtan Wang; Motonari Uesugi

doi:10.1002/anie.201410339

Live-cell imaging of endogenous mRNAs with a small molecule

Shin Ichi Sato, Mizuki Watanabe, Yousuke Katsuda, Asako Murata, Dan Ohtan Wang, Motonari Uesugi

Biology

Research output: Contribution to journal › Article › peer-review

Abstract

Determination of subcellular localization and dynamics of mRNA is increasingly important to understanding gene expression. A new convenient and versatile method is reported that permits spatiotemporal imaging of specific nonengineered RNAs in living cells. The method uses transfection of a plasmid encoding a gene-specific RNA aptamer, combined with a cell-permeable synthetic small molecule, the fluorescence of which is restored only when the RNA aptamer hybridizes with its cognitive mRNA. The method was validated by live-cell imaging of the endogenous mRNA of b-actin. Application of the technology to mRNAs of a total of 84 human cytoskeletal genes allowed us to observe cellular dynamics of several endogenous mRNAs including arfaptin-2, cortactin, and cytoplasmic FMR1-interacting protein 2. The RNA-imaging technology and its further optimization might permit live-cell imaging of any RNA molecules.

Original language	English (US)
Pages (from-to)	1855-1858
Number of pages	4
Journal	Angewandte Chemie - International Edition
Volume	54
Issue number	6
DOIs	https://doi.org/10.1002/anie.201410339
State	Published - Feb 9 2015

Keywords

Fluorescence
Live-cell imaging
RNA
RNA dynamics
Small molecules

ASJC Scopus subject areas

Catalysis
General Chemistry

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10.1002/anie.201410339

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title = "Live-cell imaging of endogenous mRNAs with a small molecule",

abstract = "Determination of subcellular localization and dynamics of mRNA is increasingly important to understanding gene expression. A new convenient and versatile method is reported that permits spatiotemporal imaging of specific nonengineered RNAs in living cells. The method uses transfection of a plasmid encoding a gene-specific RNA aptamer, combined with a cell-permeable synthetic small molecule, the fluorescence of which is restored only when the RNA aptamer hybridizes with its cognitive mRNA. The method was validated by live-cell imaging of the endogenous mRNA of b-actin. Application of the technology to mRNAs of a total of 84 human cytoskeletal genes allowed us to observe cellular dynamics of several endogenous mRNAs including arfaptin-2, cortactin, and cytoplasmic FMR1-interacting protein 2. The RNA-imaging technology and its further optimization might permit live-cell imaging of any RNA molecules.",

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AU - Sato, Shin Ichi

AU - Watanabe, Mizuki

AU - Katsuda, Yousuke

AU - Murata, Asako

AU - Wang, Dan Ohtan

AU - Uesugi, Motonari

PY - 2015/2/9

Y1 - 2015/2/9

N2 - Determination of subcellular localization and dynamics of mRNA is increasingly important to understanding gene expression. A new convenient and versatile method is reported that permits spatiotemporal imaging of specific nonengineered RNAs in living cells. The method uses transfection of a plasmid encoding a gene-specific RNA aptamer, combined with a cell-permeable synthetic small molecule, the fluorescence of which is restored only when the RNA aptamer hybridizes with its cognitive mRNA. The method was validated by live-cell imaging of the endogenous mRNA of b-actin. Application of the technology to mRNAs of a total of 84 human cytoskeletal genes allowed us to observe cellular dynamics of several endogenous mRNAs including arfaptin-2, cortactin, and cytoplasmic FMR1-interacting protein 2. The RNA-imaging technology and its further optimization might permit live-cell imaging of any RNA molecules.

AB - Determination of subcellular localization and dynamics of mRNA is increasingly important to understanding gene expression. A new convenient and versatile method is reported that permits spatiotemporal imaging of specific nonengineered RNAs in living cells. The method uses transfection of a plasmid encoding a gene-specific RNA aptamer, combined with a cell-permeable synthetic small molecule, the fluorescence of which is restored only when the RNA aptamer hybridizes with its cognitive mRNA. The method was validated by live-cell imaging of the endogenous mRNA of b-actin. Application of the technology to mRNAs of a total of 84 human cytoskeletal genes allowed us to observe cellular dynamics of several endogenous mRNAs including arfaptin-2, cortactin, and cytoplasmic FMR1-interacting protein 2. The RNA-imaging technology and its further optimization might permit live-cell imaging of any RNA molecules.

KW - Fluorescence

KW - Live-cell imaging

KW - RNA

KW - RNA dynamics

KW - Small molecules

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Live-cell imaging of endogenous mRNAs with a small molecule

Abstract

Keywords

ASJC Scopus subject areas

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