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A color-shifting near-Infrared Fluorescent Aptamer-Fluorophore Module for Live-Cell RNA Imaging

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Fluorescent light-up RNA aptamers (FLAPs) have become promising tools for visualizing RNAs in living cells. Specific binding of FLAPs to their non-fluorescent cognate ligands results in a dramatic fluorescence increase, thereby allowing RNA imaging. Here, we present a color-shifting aptamer-fluoroph...

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Bibliographic Details
Main Authors: Zhang, Jingye (Author) , Wang, Lu (Author) , Jäschke, Andres (Author) , Sünbül, Murat (Author)
Format: Article (Journal)
Language:English
Published: 20. August 2021
In: Angewandte Chemie
Year: 2021, Volume: 133, Issue: 39, Pages: 21611-21618
ISSN:1521-3757
DOI:10.1002/ange.202107250
Online Access:Verlag, kostenfrei, Volltext: https://doi.org/10.1002/ange.202107250
Verlag, kostenfrei, Volltext: https://onlinelibrary.wiley.com/doi/abs/10.1002/ange.202107250
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Author Notes:Jingye Zhang, Lu Wang, Andres Jäschke and Murat Sunbul
Description
Summary:Fluorescent light-up RNA aptamers (FLAPs) have become promising tools for visualizing RNAs in living cells. Specific binding of FLAPs to their non-fluorescent cognate ligands results in a dramatic fluorescence increase, thereby allowing RNA imaging. Here, we present a color-shifting aptamer-fluorophore system, where the free dye is cyan fluorescent and the aptamer-dye complex is near-infrared (NIR) fluorescent. Unlike other reported FLAPs, this system enables ratiometric RNA imaging. To design the color-shifting system, we synthesized a series of environmentally sensitive benzopyrylium-coumarin hybrid fluorophores which exist in equilibrium between a cyan fluorescent spirocyclic form and a NIR fluorescent zwitterionic form. As an RNA tag, we evolved a 38-nucleotide aptamer that selectively binds the zwitterionic forms with nanomolar affinity. We used this system as a light-up RNA marker to image mRNAs in the NIR region and demonstrated its utility in ratiometric analysis of target RNAs expressed at different levels in single cells.
Item Description:Gesehen am 29.03.2023
Physical Description:Online Resource
ISSN:1521-3757
DOI:10.1002/ange.202107250

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