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Issue 10, 2017
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Fluorescent turn-on probes for wash-free mRNA imaging via covalent site-specific enzymatic labeling

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Abstract

Investigating the many roles RNA plays in cellular regulation and function has increased demand for tools to explore RNA tracking and localization within cells. Our recently reported RNA-TAG (transglycosylation at guanine) approach uses an RNA-modifying enzyme, tRNA-guanine transglycosylase (TGT), to accomplish covalent labeling of an RNA of interest with fluorescent tracking agents in a highly selective and efficient manner. Unfortunately, labeling by this method currently suffers from a high nonspecific fluorescent background and is currently unsuitable for imaging RNA within complex cellular environments. Herein we report the design and synthesis of novel fluorogenic thiazole orange probes that significantly lower nonspecific binding and background fluorescence and, as a result, provide up to a 100-fold fluorescence intensity increase after labeling. Using these fluorogenic labeling agents, we were able to image mRNA expressed in Chinese Hamster Ovary cells in a wash-free manner.

Graphical abstract: Fluorescent turn-on probes for wash-free mRNA imaging via covalent site-specific enzymatic labeling

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Publication details

The article was received on 19 Jul 2017, accepted on 28 Aug 2017 and first published on 29 Aug 2017


Article type: Edge Article
DOI: 10.1039/C7SC03150E
Citation: Chem. Sci., 2017,8, 7169-7173
  • Open access: Creative Commons BY license
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    Fluorescent turn-on probes for wash-free mRNA imaging via covalent site-specific enzymatic labeling

    C. Y. Zhou, S. C. Alexander and N. K. Devaraj, Chem. Sci., 2017, 8, 7169
    DOI: 10.1039/C7SC03150E

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