Live-cell imaging of endogenous mRNAs with a small molecule

2016 Fiscal Year Final Research Report

• Project/Area Number: 26440005
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Molecular biology
• Research Institution: Kyoto University
• Principal Investigator: SATO SHINICHI 京都大学, 化学研究所, 准教授 (70534478)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: RNA生細胞内イメージング / 蛍光プローブ / ケミカルバイオロジー / RNAアプタマー

Outline of Final Research Achievements

Determination of subcellular localization and dynamics of mRNA is increasingly important to understanding gene expression. We developed a new convenient and versatile method 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 whose fluorescence 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 β-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 ARFIP2, CTTN, and CYFIP2. The RNA-imaging technology and its further optimization might permit live-cell imaging of any RNA molecules.

Free Research Field

ケミカルバイオロジー