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

Author

Listed:
  • Hela Benaissa

    (Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Laboratoire des Biomolécules, LBM
    PASTEUR, Department of Chemistry, École Normale Supérieure, Université PSL, Sorbonne Université, CNRS)

  • 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
    PASTEUR, Department of Chemistry, École Normale Supérieure, Université PSL, Sorbonne Université, CNRS
    Janelia Research Campus, Howard Hughes Medical Institute)

  • Chenge Li

    (PASTEUR, Department of Chemistry, École Normale Supérieure, Université PSL, Sorbonne Université, CNRS
    Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University
    Shanghai Cancer Institute, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University)

  • 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
    Université de Paris, Institute of Psychiatry and Neuroscience of Paris, INSERM U1266, Membrane Traffic in Healthy & Diseased Brain)

  • Nicolas Pietrancosta

    (Sorbonne Université, École Normale Supérieure, Université PSL, CNRS, Laboratoire des Biomolécules, LBM
    Neuroscience Paris Seine-Institut de Biologie Paris Seine (NPS-IBPS) INSERM, CNRS, Sorbonne Université)

  • 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
    PASTEUR, Department of Chemistry, École Normale Supérieure, Université PSL, Sorbonne Université, CNRS
    Institut Universitaire de France)

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.

Suggested Citation

  • 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, 2021. "Engineering of a fluorescent chemogenetic reporter with tunable color for advanced live-cell imaging," Nature Communications, Nature, vol. 12(1), pages 1-17, December.
    • Handle: RePEc:nat:natcom:v:12:y:2021:i:1:d:10.1038_s41467-021-27334-0
    DOI: 10.1038/s41467-021-27334-0
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    References listed on IDEAS

    as
    1. Alison G. Tebo & Arnaud Gautier, 2019. "Author Correction: A split fluorescent reporter with rapid and reversible complementation," Nature Communications, Nature, vol. 10(1), pages 1-2, December.
    2. Giulia Bertolin & Florian Sizaire & Gaëtan Herbomel & David Reboutier & Claude Prigent & Marc Tramier, 2016. "A FRET biosensor reveals spatiotemporal activation and functions of aurora kinase A in living cells," Nature Communications, Nature, vol. 7(1), pages 1-16, November.
    3. Alison G. Tebo & Arnaud Gautier, 2019. "A split fluorescent reporter with rapid and reversible complementation," Nature Communications, Nature, vol. 10(1), pages 1-8, December.
    4. Aymeric Leray & Sergi Padilla-Parra & Julien Roul & Laurent Héliot & Marc Tramier, 2013. "827Spatio-Temporal Quantification of FRET in Living Cells by Fast Time-Domain FLIM: A Comparative Study of Non-Fitting Methods," PLOS ONE, Public Library of Science, vol. 8(7), pages 1-16, July.
    5. Andreas Crameri & Sun-Ai Raillard & Ericka Bermudez & Willem P. C. Stemmer, 1998. "DNA shuffling of a family of genes from diverse species accelerates directed evolution," Nature, Nature, vol. 391(6664), pages 288-291, January.
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