Fast-exchanging spirocyclic rhodamine probes for aptamer-based super-resolution RNA imaging.


Journal

Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555

Informations de publication

Date de publication:
30 06 2023
Historique:
received: 04 11 2022
accepted: 22 06 2023
medline: 3 7 2023
pubmed: 1 7 2023
entrez: 30 6 2023
Statut: epublish

Résumé

Live-cell RNA imaging with high spatial and temporal resolution remains a major challenge. Here we report the development of RhoBAST:SpyRho, a fluorescent light-up aptamer (FLAP) system ideally suited for visualizing RNAs in live or fixed cells with various advanced fluorescence microscopy modalities. Overcoming problems associated with low cell permeability, brightness, fluorogenicity, and signal-to-background ratio of previous fluorophores, we design a novel probe, SpyRho (Spirocyclic Rhodamine), which tightly binds to the RhoBAST aptamer. High brightness and fluorogenicity is achieved by shifting the equilibrium between spirolactam and quinoid. With its high affinity and fast ligand exchange, RhoBAST:SpyRho is a superb system for both super-resolution SMLM and STED imaging. Its excellent performance in SMLM and the first reported super-resolved STED images of specifically labeled RNA in live mammalian cells represent significant advances over other FLAPs. The versatility of RhoBAST:SpyRho is further demonstrated by imaging endogenous chromosomal loci and proteins.

Identifiants

pubmed: 37391423
doi: 10.1038/s41467-023-39611-1
pii: 10.1038/s41467-023-39611-1
pmc: PMC10313827
doi:

Substances chimiques

Rhodamines 0
Fluorescent Dyes 0
Ionophores 0
Oligonucleotides 0
RNA 63231-63-0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

3879

Informations de copyright

© 2023. The Author(s).

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Auteurs

Daniel Englert (D)

Institute of Pharmacy and Molecular Biotechnology (IPMB), Heidelberg University, Heidelberg, Germany.

Eva-Maria Burger (EM)

Institute of Pharmacy and Molecular Biotechnology (IPMB), Heidelberg University, Heidelberg, Germany.

Franziska Grün (F)

Institute of Pharmacy and Molecular Biotechnology (IPMB), Heidelberg University, Heidelberg, Germany.

Mrigank S Verma (MS)

Institute of Applied Physics (APH), Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany.

Jens Lackner (J)

Institute of Applied Physics (APH), Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany.

Marko Lampe (M)

Advanced Light Microscopy Facility, European Molecular Biology Laboratory, Heidelberg, Germany.

Bastian Bühler (B)

Institute of Pharmacy and Molecular Biotechnology (IPMB), Heidelberg University, Heidelberg, Germany.

Janin Schokolowski (J)

Institute of Pharmacy and Molecular Biotechnology (IPMB), Heidelberg University, Heidelberg, Germany.

G Ulrich Nienhaus (GU)

Institute of Applied Physics (APH), Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany. uli@uiuc.edu.
Institute of Nanotechnology (INT), Karlsruhe Institute of Technology (KIT), Eggenstein-Leopoldshafen, Germany. uli@uiuc.edu.
Institute of Biological and Chemical Systems (IBCS), Karlsruhe Institute of Technology (KIT), Eggenstein-Leopoldshafen, Germany. uli@uiuc.edu.
Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL, USA. uli@uiuc.edu.

Andres Jäschke (A)

Institute of Pharmacy and Molecular Biotechnology (IPMB), Heidelberg University, Heidelberg, Germany. jaeschke@uni-hd.de.

Murat Sunbul (M)

Institute of Pharmacy and Molecular Biotechnology (IPMB), Heidelberg University, Heidelberg, Germany. msunbul@uni-heidelberg.de.

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Classifications MeSH