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

Alexis Autour, Sunny Jeng, Adam Cawte, Amir Abdolahzadeh, Angela Galli, Shanker S. S. Panchapakesan, David Rueda (), Michael Ryckelynck () and Peter J. Unrau ()
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Alexis Autour: Université de Strasbourg, CNRS, Architecture et Réactivité de l’ARN, UPR 9002
Sunny Jeng: Simon Fraser University
Adam Cawte: MRC London Institute of Medical Sciences
Amir Abdolahzadeh: Simon Fraser University
Angela Galli: Simon Fraser University
Shanker S. S. Panchapakesan: Simon Fraser University
David Rueda: MRC London Institute of Medical Sciences
Michael Ryckelynck: Université de Strasbourg, CNRS, Architecture et Réactivité de l’ARN, UPR 9002
Peter J. Unrau: Simon Fraser University

Nature Communications, 2018, vol. 9, issue 1, 1-12

Abstract: 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.

Date: 2018
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DOI: 10.1038/s41467-018-02993-8

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