A tunable and versatile chemogenetic near-infrared fluorescent reporter

Abstract Near-infrared (NIR) fluorescent reporters open interesting perspectives for multiplexed imaging with higher contrast and depth using less toxic light. Here, we propose nirFAST, a small (14 kDa) chemogenetic NIR fluorescent reporter, displaying higher cellular brightness compared to top-perf...

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Main Authors: Lina El Hajji, Benjamin Bunel, Octave Joliot, Chenge Li, Alison G. Tebo, Christine Rampon, Michel Volovitch, Evelyne Fischer, Nicolas Pietrancosta, Franck Perez, Xavier Morin, Sophie Vriz, Arnaud Gautier
Format: Article
Language:English
Published: Nature Portfolio 2025-03-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-58017-9
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author Lina El Hajji
Benjamin Bunel
Octave Joliot
Chenge Li
Alison G. Tebo
Christine Rampon
Michel Volovitch
Evelyne Fischer
Nicolas Pietrancosta
Franck Perez
Xavier Morin
Sophie Vriz
Arnaud Gautier
author_facet Lina El Hajji
Benjamin Bunel
Octave Joliot
Chenge Li
Alison G. Tebo
Christine Rampon
Michel Volovitch
Evelyne Fischer
Nicolas Pietrancosta
Franck Perez
Xavier Morin
Sophie Vriz
Arnaud Gautier
author_sort Lina El Hajji
collection DOAJ
description Abstract Near-infrared (NIR) fluorescent reporters open interesting perspectives for multiplexed imaging with higher contrast and depth using less toxic light. Here, we propose nirFAST, a small (14 kDa) chemogenetic NIR fluorescent reporter, displaying higher cellular brightness compared to top-performing NIR fluorescent proteins. nirFAST binds and stabilizes the fluorescent state of synthetic cell permeant fluorogenic chromophores (so-called fluorogens), otherwise dark when free. nirFAST displays tunable NIR, far-red or red emission through change of fluorogen. nirFAST allows imaging and spectral multiplexing in live cultured mammalian cells, chicken embryo tissues and zebrafish larvae. Its suitability for stimulated emission depletion nanoscopy enabled protein imaging with subdiffraction resolution in live cells. nirFAST enabled the design of a two-color cell cycle indicator for monitoring the different phases of the cell cycle. Finally, bisection of nirFAST allowed the design of a chemically induced dimerization technology with NIR fluorescence readout, enabling the control and visualization of protein proximity.
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issn 2041-1723
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spelling doaj-art-578f279bcc6d4ac59b6b7750b56590e42025-08-20T02:56:16ZengNature PortfolioNature Communications2041-17232025-03-0116111710.1038/s41467-025-58017-9A tunable and versatile chemogenetic near-infrared fluorescent reporterLina El Hajji0Benjamin Bunel1Octave Joliot2Chenge Li3Alison G. Tebo4Christine Rampon5Michel Volovitch6Evelyne Fischer7Nicolas Pietrancosta8Franck Perez9Xavier Morin10Sophie Vriz11Arnaud Gautier12Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Chimie Physique et Chimie du Vivant (CPCV)Institut de Biologie de l’ENS (IBENS), École Normale Supérieure, CNRS, INSERM, Université PSLInstitut Curie, Université PSL, CNRS UMR144Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Chimie Physique et Chimie du Vivant (CPCV)Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Chimie Physique et Chimie du Vivant (CPCV)Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Chimie Physique et Chimie du Vivant (CPCV)Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Chimie Physique et Chimie du Vivant (CPCV)Institut de Biologie de l’ENS (IBENS), École Normale Supérieure, CNRS, INSERM, Université PSLSorbonne Université, École Normale Supérieure, Université PSL, CNRS, Chimie Physique et Chimie du Vivant (CPCV)Institut Curie, Université PSL, CNRS UMR144Institut de Biologie de l’ENS (IBENS), École Normale Supérieure, CNRS, INSERM, Université PSLSorbonne Université, École Normale Supérieure, Université PSL, CNRS, Chimie Physique et Chimie du Vivant (CPCV)Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Chimie Physique et Chimie du Vivant (CPCV)Abstract Near-infrared (NIR) fluorescent reporters open interesting perspectives for multiplexed imaging with higher contrast and depth using less toxic light. Here, we propose nirFAST, a small (14 kDa) chemogenetic NIR fluorescent reporter, displaying higher cellular brightness compared to top-performing NIR fluorescent proteins. nirFAST binds and stabilizes the fluorescent state of synthetic cell permeant fluorogenic chromophores (so-called fluorogens), otherwise dark when free. nirFAST displays tunable NIR, far-red or red emission through change of fluorogen. nirFAST allows imaging and spectral multiplexing in live cultured mammalian cells, chicken embryo tissues and zebrafish larvae. Its suitability for stimulated emission depletion nanoscopy enabled protein imaging with subdiffraction resolution in live cells. nirFAST enabled the design of a two-color cell cycle indicator for monitoring the different phases of the cell cycle. Finally, bisection of nirFAST allowed the design of a chemically induced dimerization technology with NIR fluorescence readout, enabling the control and visualization of protein proximity.https://doi.org/10.1038/s41467-025-58017-9
spellingShingle Lina El Hajji
Benjamin Bunel
Octave Joliot
Chenge Li
Alison G. Tebo
Christine Rampon
Michel Volovitch
Evelyne Fischer
Nicolas Pietrancosta
Franck Perez
Xavier Morin
Sophie Vriz
Arnaud Gautier
A tunable and versatile chemogenetic near-infrared fluorescent reporter
Nature Communications
title A tunable and versatile chemogenetic near-infrared fluorescent reporter
title_full A tunable and versatile chemogenetic near-infrared fluorescent reporter
title_fullStr A tunable and versatile chemogenetic near-infrared fluorescent reporter
title_full_unstemmed A tunable and versatile chemogenetic near-infrared fluorescent reporter
title_short A tunable and versatile chemogenetic near-infrared fluorescent reporter
title_sort tunable and versatile chemogenetic near infrared fluorescent reporter
url https://doi.org/10.1038/s41467-025-58017-9
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