Live‐Cell Imaging of Endogenous mRNAs with a Small Molecule |
| |
Authors: | Dr Shin‐ichi Sato Dr Mizuki Watanabe Dr Yousuke Katsuda Dr Asako Murata Dr Dan Ohtan Wang Prof Motonari Uesugi |
| |
Institution: | Institute for Integrated Cell‐Material Sciences (WPI‐iCeMS) and Institute for Chemical Research, Kyoto University, Uji, Kyoto 611‐0011 (Japan) |
| |
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 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 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 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. |
| |
Keywords: | fluorescence live‐cell imaging RNA RNA dynamics small molecules |
|
|