Engineering of a fluorescent chemogenetic reporter with tunable color for advanced live-cell imaging

Hela Benaissa, Karim Ounoughi, Isabelle Aujard, Evelyne Fischer, Rosette Goïame, Julie Nguyen, Alison G. Tebo, Chenge Li, Thomas Saux, Giulia Bertolin, Marc Tramier, Lydia Danglot, Nicolas Pietrancosta, Xavier Morin, Ludovic Jullien and Arnaud Gautier ()
Additional contact information
Hela Benaissa: Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Laboratoire des Biomolécules, LBM
Karim Ounoughi: PASTEUR, Department of Chemistry, École Normale Supérieure, Université PSL, Sorbonne Université, CNRS
Isabelle Aujard: PASTEUR, Department of Chemistry, École Normale Supérieure, Université PSL, Sorbonne Université, CNRS
Evelyne Fischer: Institut de Biologie de l’ENS (IBENS), École Normale Supérieure, CNRS, INSERM, Université PSL
Rosette Goïame: Institut de Biologie de l’ENS (IBENS), École Normale Supérieure, CNRS, INSERM, Université PSL
Julie Nguyen: Université de Paris, NeurImag Imaging Facility, Institute of Psychiatry and Neuroscience of Paris, INSERM U1266
Alison G. Tebo: Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Laboratoire des Biomolécules, LBM
Chenge Li: PASTEUR, Department of Chemistry, École Normale Supérieure, Université PSL, Sorbonne Université, CNRS
Thomas Saux: PASTEUR, Department of Chemistry, École Normale Supérieure, Université PSL, Sorbonne Université, CNRS
Giulia Bertolin: University of Rennes, Centre National de la Recherche Scientifique (CNRS), (IGDR) Institute of Genetics and Development of Rennes, Unité Mixte de Recherche (UMR) 6290
Marc Tramier: University of Rennes, Centre National de la Recherche Scientifique (CNRS), (IGDR) Institute of Genetics and Development of Rennes, Unité Mixte de Recherche (UMR) 6290
Lydia Danglot: Université de Paris, NeurImag Imaging Facility, Institute of Psychiatry and Neuroscience of Paris, INSERM U1266
Nicolas Pietrancosta: Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Laboratoire des Biomolécules, LBM
Xavier Morin: Institut de Biologie de l’ENS (IBENS), École Normale Supérieure, CNRS, INSERM, Université PSL
Ludovic Jullien: PASTEUR, Department of Chemistry, École Normale Supérieure, Université PSL, Sorbonne Université, CNRS
Arnaud Gautier: Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Laboratoire des Biomolécules, LBM

Nature Communications, 2021, vol. 12, issue 1, 1-17

Abstract: Abstract Biocompatible fluorescent reporters with spectral properties spanning the entire visible spectrum are indispensable tools for imaging the biochemistry of living cells and organisms in real time. Here, we report the engineering of a fluorescent chemogenetic reporter with tunable optical and spectral properties. A collection of fluorogenic chromophores with various electronic properties enables to generate bimolecular fluorescent assemblies that cover the visible spectrum from blue to red using a single protein tag engineered and optimized by directed evolution and rational design. The ability to tune the fluorescence color and properties through simple molecular modulation provides a broad experimental versatility for imaging proteins in live cells, including neurons, and in multicellular organisms, and opens avenues for optimizing Förster resonance energy transfer (FRET) biosensors in live cells. The ability to tune the spectral properties and fluorescence performance enables furthermore to match the specifications and requirements of advanced super-resolution imaging techniques.

Date: 2021
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DOI: 10.1038/s41467-021-27334-0

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