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 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.
authors.affiliation: Institute for Integrated Cell-Material Sciences (WPI-iCeMS) and Institute for Chemical Research, Kyoto University, Uji, Kyoto 611-0011 (Japan). ssato@scl.kyoto-u.ac.jp.
authors.forename: Shin-ichi
authors.initials: S
authors.lastname: Sato
authors.forename: Mizuki
authors.initials: M
authors.lastname: Watanabe
authors.forename: Yousuke
authors.initials: Y
authors.lastname: Katsuda
authors.forename: Asako
authors.initials: A
authors.lastname: Murata
authors.forename: Dan Ohtan
authors.initials: DO
authors.lastname: Wang
authors.forename: Motonari
authors.initials: M
authors.lastname: Uesugi
mesh_terms: D001483:Base Sequence; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics*
chemical_list: D012333:RNA, Messenger
keywords: RNA; RNA dynamics; fluorescence; live-cell imaging; small molecules