Illuminating biology with fluorescent RNAs

Murat Sünbül

Heidelberg Universität, Institute of Pharmacy and Molecular Biotechnology, Im Neuenheimer Feld 364, 69120 Heidelberg [DE], msunbul@uni-heidelberg.de

Author(s):
Murat Sünbül

We have recently developed RhoBAST1, a genetically encoded fluorescent RNA tag that binds fluorogenic rhodamine dyes2 with exceptionally rapid association and dissociation kinetics. These quick binding and unbinding events cause intermittent fluorescence emission (blinking), facilitating super-resolution imaging of RNAs tagged with RhoBAST by single-molecule localization microscopy (SMLM).1,2 Furthermore, the fast and continuous dye exchange is particularly beneficial for STED microscopy, which commonly faces issues with fluorophore photobleaching.2 The rapid exchange kinetics allow bleached dyes to be replaced by fresh ones within seconds (or less), thereby restoring the fluorescence signal.

Here, I will demonstrate the remarkable potential and versatility of RhoBAST technology for imaging and tracking3 not only RNA but also DNA and proteins using various super-resolution imaging techniques.4

[1] Sunbul et al. “Super-resolution RNA imaging using a rhodamine-binding aptamer with fast exchange kinetics” Nature Biotechnology, 2021, 39, 686

[2] Englert et al. “Fast-exchanging spirocyclic rhodamine probes for aptamer-based super-resolution RNA imaging” Nature Communications, 2023, 14, 3879

[3] Bühler et al. “Avidity-based bright and photostable light-up aptamers for single-molecule mRNA imaging” Nature Chemical Biology, 2023, 19, 478

[4] Grün et al. “Super-resolved protein imaging using bifunctional light-up aptamers” bioRxiv, 2024, 2024.01.26.577321

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