Journal
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
Volume 54, Issue 6, Pages 1855-1858Publisher
WILEY-V C H VERLAG GMBH
DOI: 10.1002/anie.201410339
Keywords
fluorescence; live-cell imaging; RNA; RNA dynamics; small molecules
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Funding
- JSPS [26220206, 26440005]
- JST [AS242Z01268P]
- ZE Research Program, IAE [B-8]
- iCeMS research acceleration grant
- World Premier International Research Center Initiative (WPI), MEXT (Japan)
- New Energy and Industrial Technology Development Organization (NEDO) of Japan
- Yokogawa Electric Corporation
- Grants-in-Aid for Scientific Research [26560445, 26220206, 26350972, 26702038, 26102728, 26440005] Funding Source: KAKEN
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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 non-engineered 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 -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 protein2. The RNA-imaging technology and its further optimization might permit live-cell imaging of any RNA molecules.
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