Kinetic and Mechanistic Characterization of RSpFAST, a Non-Covalent Versatile Chemogenetic Platform for Fluorescence Imaging
Abstract
Reversibly photoswitchable fluorophores have enabled a broad range of applications in advanced fluorescence bioimaging. Here, we introduce RSpFAST, a new class of reversibly photoswitchable fluorescent labels that combine a biomolecular host (pFAST protein tag) with a reversibly photoisomerizable guest (fluorogen), allowing fluorescence brightness to be modulated through illumination and molecular complexation. We combine thermokinetic, photochemical, and structural investigations to obtain a comprehensive mechanistic and kinetic understanding of RSpFAST. Building on this theoretical framework, we demonstrate in both live and fixed cells that RSpFAST exhibits an unprecedented dual behavior: a stable and wash-free fluorescent labeling tag turns into a negative reversible photoswitcher by decreasing the fluorogen concentration and increasing light intensity. In this photoejection-driven kinetic regime, RSpFAST is shown to be an efficient marker for dynamic contrast and super-resolution microscopy.
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BibTeX
@article{shpinov2026kinetic,
title = {{Kinetic and Mechanistic Characterization of RSpFAST, a Non-Covalent Versatile Chemogenetic Platform for Fluorescence Imaging}},
author = {Yuriy Shpinov and Mrinal Mandal and Vincent van Deuren and Alienor Lahlou and Matthias Le Bec and Raja Chouket and Chaima Hadj Moussa and Clara Bonin and Hessam Sepasi Tehrani and Ian Coghill and Lina El Hajji and Karim Ounoughi and Jaime Franco Pinto and Marie-Aude Plamont and Philippe Pelupessy and Isabel Ayala and Franck Perez and Isabelle Aujard and Thomas Le Saux and Arnaud Gautier and Peter Dedecker and Bernhard Brutscher and Ludovic Jullien},
journal = {Journal of the American Chemical Society},
year = {2026},
month = aug,
doi = {10.1021/jacs.6c08277},
url = {https://hal.science/hal-05803513v1},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Kinetic and Mechanistic Characterization of RSpFAST, a Non-Covalent Versatile Chemogenetic Platform for Fluorescence Imaging
AU - Yuriy Shpinov
AU - Mrinal Mandal
AU - Vincent van Deuren
AU - Alienor Lahlou
AU - Matthias Le Bec
AU - Raja Chouket
AU - Chaima Hadj Moussa
AU - Clara Bonin
AU - Hessam Sepasi Tehrani
AU - Ian Coghill
AU - Lina El Hajji
AU - Karim Ounoughi
AU - Jaime Franco Pinto
AU - Marie-Aude Plamont
AU - Philippe Pelupessy
AU - Isabel Ayala
AU - Franck Perez
AU - Isabelle Aujard
AU - Thomas Le Saux
AU - Arnaud Gautier
AU - Peter Dedecker
AU - Bernhard Brutscher
AU - Ludovic Jullien
PY - 2026
DA - 2026/08/25
JO - Journal of the American Chemical Society
DO - 10.1021/jacs.6c08277
UR - https://hal.science/hal-05803513v1
AB - Reversibly photoswitchable fluorophores have enabled a broad range of applications in advanced fluorescence bioimaging. Here, we introduce RSpFAST, a new class of reversibly photoswitchable fluorescent labels that combine a biomolecular host (pFAST protein tag) with a reversibly photoisomerizable guest (fluorogen), allowing fluorescence brightness to be modulated through illumination and molecular complexation. We combine thermokinetic, photochemical, and structural investigations to obtain a comprehensive mechanistic and kinetic understanding of RSpFAST. Building on this theoretical framework, we demonstrate in both live and fixed cells that RSpFAST exhibits an unprecedented dual behavior: a stable and wash-free fluorescent labeling tag turns into a negative reversible photoswitcher by decreasing the fluorogen concentration and increasing light intensity. In this photoejection-driven kinetic regime, RSpFAST is shown to be an efficient marker for dynamic contrast and super-resolution microscopy.
SN - 0002-7863
SN - 1520-5126
ER -