A color-shifting near-infrared fluorescent aptamer-fluorophore module for live-cell RNA imaging

• Verfasst von: Zhang, Jingye [VerfasserIn]
• Verfasst von: Wang, Lu [VerfasserIn]
• Verfasst von: Jäschke, Andres [VerfasserIn]
• Verfasst von: Sünbül, Murat [VerfasserIn]
• Titel: A color-shifting near-infrared fluorescent aptamer-fluorophore module for live-cell RNA imaging
• Verf.angabe: Jingye Zhang, Lu Wang, Andres Jäschke, and Murat Sunbul
• E-Jahr: 2021
• Jahr: August 20, 2021
• Umfang: 8 S.
• Fussnoten: Gesehen am 29.03.2023
• Titel Quelle: Enthalten in: Angewandte Chemie. International edition
• Ort Quelle: Weinheim : Wiley-VCH, 1998
• Jahr Quelle: 2021
• Band/Heft Quelle: 60(2021), 39, Seite 21441-21448
• ISSN Quelle: 1521-3773
• DOI: doi:10.1002/anie.202107250
• Datenträger: Online-Ressource
• Sprache: eng
• Sach-SW: fluorophores
• Sach-SW: ratiometric imaging
• Sach-SW: RNA aptamer
• Sach-SW: RNA imaging
• Sach-SW: SELEX
• K10plus-PPN: 1840449748
• K10plus-PPN:
  Universitätsbibliothek
• Lokale URL UB: Zum Volltext

Abstract

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.