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Nat Commun. 2018 Feb 13;9(1):656. doi: 10.1038/s41467-018-02993-8.

Fluorogenic RNA Mango aptamers for imaging small non-coding RNAs in mammalian cells.

Author information

1
Université de Strasbourg, CNRS, Architecture et Réactivité de l'ARN, UPR 9002, 67000, Strasbourg, France.
2
Department of Molecular Biology and Biochemistry, Simon Fraser University, 8888 University Drive, Burnaby, BC, V5A 1S6, Canada.
3
Single Molecule Imaging Group, MRC London Institute of Medical Sciences, Du Cane Road, London, W12 0NN, UK.
4
Department of Medicine, Molecular Virology, Imperial College London, Du Cane Road, London, W12 0NN, UK.
5
Single Molecule Imaging Group, MRC London Institute of Medical Sciences, Du Cane Road, London, W12 0NN, UK. david.rueda@imperial.ac.uk.
6
Department of Medicine, Molecular Virology, Imperial College London, Du Cane Road, London, W12 0NN, UK. david.rueda@imperial.ac.uk.
7
Université de Strasbourg, CNRS, Architecture et Réactivité de l'ARN, UPR 9002, 67000, Strasbourg, France. m.ryckelynck@unistra.fr.
8
Department of Molecular Biology and Biochemistry, Simon Fraser University, 8888 University Drive, Burnaby, BC, V5A 1S6, Canada. punrau@sfu.ca.

Abstract

Despite having many key roles in cellular biology, directly imaging biologically important RNAs has been hindered by a lack of fluorescent tools equivalent to the fluorescent proteins available to study cellular proteins. Ideal RNA labelling systems must preserve biological function, have photophysical properties similar to existing fluorescent proteins, and be compatible with established live and fixed cell protein labelling strategies. Here, we report a microfluidics-based selection of three new high-affinity RNA Mango fluorogenic aptamers. Two of these are as bright or brighter than enhanced GFP when bound to TO1-Biotin. Furthermore, we show that the new Mangos can accurately image the subcellular localization of three small non-coding RNAs (5S, U6, and a box C/D scaRNA) in fixed and live mammalian cells. These new aptamers have many potential applications to study RNA function and dynamics both in vitro and in mammalian cells.

PMID:
29440634
PMCID:
PMC5811451
DOI:
10.1038/s41467-018-02993-8
[Indexed for MEDLINE]
Free PMC Article

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