A single-chain and fast-responding light-inducible Cre recombinase as a novel optogenetic switch

  1. Hélène Duplus-Bottin
  2. Martin Spichty
  3. Gérard Triqueneaux
  4. Christophe Place
  5. Philippe Emmanuel Mangeot
  6. Théophile Ohlmann
  7. Franck Vittoz
  8. Gaël Yvert  Is a corresponding author
  1. CNRS and Ecole Normale Superieure de Lyon, France
  2. INSERM and Ecole Normale Superieure de Lyon, France

Abstract

Optogenetics enables genome manipulations with high spatiotemporal resolution, opening exciting possibilities for fundamental and applied biological research. Here, we report the development of LiCre, a novel light-inducible Cre recombinase. LiCre is made of a single flavin-containing protein comprising the AsLOV2 photoreceptor domain of Avena sativa fused to a Cre variant carrying destabilizing mutations in its N-terminal and C-terminal domains. LiCre can be activated within minutes of illumination with blue light, without the need of additional chemicals. When compared to existing photoactivatable Cre recombinases based on two split units, LiCre displayed faster and stronger activation by light as well as a lower residual activity in the dark. LiCre was efficient both in yeast, where it allowed us to control the production of β-carotene with light, and in human cells. Given its simplicity and performances, LiCre is particularly suited for fundamental and biomedical research, as well as for controlling industrial bioprocesses.

Data availability

Raw flow-cytometry data have been deposited in Biostudies under accession code S-BSST580. Processed data used for figures are included in the supporting files.

Article and author information

Author details

  1. Hélène Duplus-Bottin

    Laboratory of Biology and Modeling of the Cell, CNRS UMR5239, CNRS and Ecole Normale Superieure de Lyon, Lyon, France
    Competing interests
    Hélène Duplus-Bottin, A patent application covering LiCre and its potential applications has been filed. Ref: FR3079832 A1 and WO2019193205. Patent applicant: CNRS; inventors: Hélène Duplus-Bottin, Martin Spichty and Gaël Yvert..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-2029-5646
  2. Martin Spichty

    Laboratory of Biology and Modeling of the Cell, CNRS UMR5239, CNRS and Ecole Normale Superieure de Lyon, Lyon, France
    Competing interests
    Martin Spichty, A patent application covering LiCre and its potential applications has been filed. Ref: FR3079832 A1 and WO2019193205. Patent applicant: CNRS; inventors: Hélène Duplus-Bottin, Martin Spichty and Gaël Yvert..
  3. Gérard Triqueneaux

    Laboratory of Biology and Modeling of the Cell, CNRS UMR5239, CNRS and Ecole Normale Superieure de Lyon, Lyon, France
    Competing interests
    No competing interests declared.
  4. Christophe Place

    Laboratory of Physics, CNRS UMR5672, CNRS and Ecole Normale Superieure de Lyon, Lyon, France
    Competing interests
    No competing interests declared.
  5. Philippe Emmanuel Mangeot

    CIRI-Centre International de Recherche en Infectiologie, Inserm U1111, CNRS UMR5308, INSERM and Ecole Normale Superieure de Lyon, Lyon, France
    Competing interests
    No competing interests declared.
  6. Théophile Ohlmann

    CIRI-Centre International de Recherche en Infectiologie, Inserm U1111, CNRS UMR5308, INSERM and Ecole Normale Superieure de Lyon, Lyon, France
    Competing interests
    No competing interests declared.
  7. Franck Vittoz

    Laboratory of Physics, CNRS UMR5672, CNRS and Ecole Normale Superieure de Lyon, Lyon, France
    Competing interests
    No competing interests declared.
  8. Gaël Yvert

    Laboratory of Biology and Modeling of the Cell, CNRS UMR5239, CNRS and Ecole Normale Superieure de Lyon, Lyon, France
    For correspondence
    Gael.Yvert@ens-lyon.fr
    Competing interests
    Gaël Yvert, A patent application covering LiCre and its potential applications has been filed. Ref: FR3079832 A1 and WO2019193205. Patent applicant: CNRS; inventors: Hélène Duplus-Bottin, Martin Spichty and Gaël Yvert..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1955-4786

Funding

H2020 European Research Council (StG-281359 (SiGHT))

  • Gaël Yvert

Centre National de la Recherche Scientifique (MITI 80 Prime READGEN)

  • Gaël Yvert

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Reviewing Editor

  1. Kevin H Gardner, CUNY Advanced Science Research Center, United States

Publication history

  1. Received: July 20, 2020
  2. Accepted: February 22, 2021
  3. Accepted Manuscript published: February 23, 2021 (version 1)
  4. Version of Record published: March 26, 2021 (version 2)

Copyright

© 2021, Duplus-Bottin et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

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  1. Hélène Duplus-Bottin
  2. Martin Spichty
  3. Gérard Triqueneaux
  4. Christophe Place
  5. Philippe Emmanuel Mangeot
  6. Théophile Ohlmann
  7. Franck Vittoz
  8. Gaël Yvert
(2021)
A single-chain and fast-responding light-inducible Cre recombinase as a novel optogenetic switch
eLife 10:e61268.
https://doi.org/10.7554/eLife.61268

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