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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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Hauptverfasser: Zhang, Jingye (VerfasserIn) , Wang, Lu (VerfasserIn) , Jäschke, Andres (VerfasserIn) , Sünbül, Murat (VerfasserIn)
Dokumenttyp: Article (Journal)
Sprache:Englisch
Veröffentlicht: August 20, 2021
In: Angewandte Chemie. International edition
Year: 2021, Jahrgang: 60, Heft: 39, Pages: 21441-21448
ISSN:1521-3773
DOI:10.1002/anie.202107250
Online-Zugang:Verlag, kostenfrei, Volltext: https://doi.org/10.1002/anie.202107250
Verlag, kostenfrei, Volltext: https://onlinelibrary.wiley.com/doi/abs/10.1002/anie.202107250
Volltext
Verfasserangaben:Jingye Zhang, Lu Wang, Andres Jäschke, and Murat Sunbul
Beschreibung
Zusammenfassung: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.
Beschreibung:Gesehen am 29.03.2023
Beschreibung:Online Resource
ISSN:1521-3773
DOI:10.1002/anie.202107250

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