A modular scaffold for cellularly-retained fluorogenic probes for sensitive cell-resolved bioactivity imaging

Publikation: Beitrag in Fachzeitschrift › Forschungsartikel › Beigetragen › Begutachtung

Beitragende

  • Philipp Mauker - , Professur für Organische Chemie (II) (OC2), Technische Universität Dresden, Ludwig-Maximilians-Universität München (LMU) (Autor:in)
  • Lucas Dessen-Weissenhorn - , Ludwig-Maximilians-Universität München (LMU) (Autor:in)
  • Carmen Zecha - , Ludwig-Maximilians-Universität München (LMU) (Autor:in)
  • Nynke A. Vepřek - , Ludwig-Maximilians-Universität München (LMU) (Autor:in)
  • Julia I. Brandmeier - , Ludwig-Maximilians-Universität München (LMU) (Autor:in)
  • Daniela Beckmann - , Klinikum der Ludwig-Maximilians-Universität (LMU) München, Ludwig-Maximilians-Universität München (LMU) (Autor:in)
  • Annabel Kitowski - , Ludwig-Maximilians-Universität München (LMU) (Autor:in)
  • Tobias Kernmayr - , Ludwig-Maximilians-Universität München (LMU) (Autor:in)
  • Julia Thorn-Seshold - , Hochschulmedizin (Medizinische Fakultät und Universitätsklinikum), Professur für Organische Chemie (II) (OC2), Institut für Klinische Chemie und Laboratoriumsmedizin, Technische Universität Dresden, Universitätsklinikum Carl Gustav Carus Dresden (Autor:in)
  • Martin Kerschensteiner - , Klinikum der Ludwig-Maximilians-Universität (LMU) München, Ludwig-Maximilians-Universität München (LMU), Munich Cluster for Systems Neurology (SyNergy) (Autor:in)
  • Oliver Thorn-Seshold - , Professur für Organische Chemie (II) (OC2) (Autor:in)

Abstract

Here, we develop a general design for high-quality fluorogenic activity probes to quantify biochemical processes within live cells, via the release of a fully cell-retained, bright fluorescent soluble product upon reaction. Live cell probes must be membrane-permeable to access intracellular biochemistry, but often that means their fluorophore products are similarly permeable resulting in rapid signal loss from the activating cell, which limits their cell-by-cell resolution as well as their sensitivity for quantifying low-turnover processes. Current strategies to retain fluorescent products within cells usually disrupt native biology e.g. by non-specific alkylation or solid precipitation. Here, scanning charge- and polarity-based approaches to swap from permeable to cell-retained states, we developed a bright fluorogenic rhodol-based platform, Trappable Green (TraG), balancing all key requirements for signal integration (rapid probe entry, but effective product retention, across many cell lines) and being modular so it can be adapted to quantify many biochemical target types (examples shown here include probes for GSH, TrxR, and H2 O2). The simple and rugged TraG scaffold can now permit straightforward implementation in a range of cell-retained enzyme activity probes, which will enable more accurate cell-resolved imaging as well as higher-sensitivity integration of low-turnover processes, without the drawbacks of alkylation or precipitation-based strategies.

Details

OriginalspracheEnglisch
Seiten (von - bis)22630-22637
Seitenumfang8
FachzeitschriftChemical science
Jahrgang16
Ausgabenummer47
PublikationsstatusVeröffentlicht - 21 Dez. 2025
Peer-Review-StatusJa

Externe IDs

WOS 001603074600001
Scopus 105023953229
ORCID /0000-0003-3981-651X/work/233047354

Schlagworte

ASJC Scopus Sachgebiete