Precise base editing for the in vivo study of developmental signaling and human pathologies in zebrafish

  1. Marion Rosello
  2. Juliette Vougny
  3. François Czarny
  4. Marina C Mione
  5. Jean-Paul Concordet
  6. Shahad Albadri  Is a corresponding author
  7. Filippo Del Bene  Is a corresponding author
  1. Institut de la Vision, France
  2. Institut Curie, France
  3. University of Trento, Italy
  4. Muséum National d'Histoire Naturelle, France

Abstract

While zebrafish is emerging as a new model system to study human diseases, an efficient methodology to generate precise point mutations at high efficiency is still lacking. Here we show that base editors can generate C-to-T point mutations with high efficiencies without other unwanted on-target mutations. In addition, we established a new editor variant recognizing an NAA PAM, expanding the base editing possibilities in zebrafish. Using these approaches, we first generated a base change in the ctnnb1 gene, mimicking oncogenic mutations of the human gene known to result in constitutive activation of endogenous Wnt signaling. Additionally, we precisely targeted several cancer-associated genes including cbl. With this last target we created a new zebrafish dwarfism model. Together our findings expand the potential of zebrafish as a model system allowing new approaches for the endogenous modulation of cell signaling pathways and the generation of precise models of human genetic disease associated-mutations.

Data availability

All data generated or analysed during this study are included in the manuscript and supporting files.

Article and author information

Author details

  1. Marion Rosello

    Developmental Biology, Institut de la Vision, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  2. Juliette Vougny

    BDD, Institut Curie, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-7361-8405
  3. François Czarny

    Developmental Biology, Institut de la Vision, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  4. Marina C Mione

    Centre for Integrative Biology (CIBIO), University of Trento, Trento, Italy
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9040-3705
  5. Jean-Paul Concordet

    Muséum National d'Histoire naturelle, Inserm U 1154, CNRS, UMR 7196, Muséum National d'Histoire Naturelle, Paris, France
    Competing interests
    The authors declare that no competing interests exist.
  6. Shahad Albadri

    Developmental Biology, Institut de la Vision, Paris, France
    For correspondence
    shahad.albadri@inserm.fr
    Competing interests
    The authors declare that no competing interests exist.
  7. Filippo Del Bene

    Developmental Biology, Institut de la Vision, Paris, France
    For correspondence
    filippo.del-bene@inserm.fr
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-8551-2846

Funding

Agence Nationale de la Recherche (ANR-18-CE16 iReelAx"")

  • Filippo Del Bene

Agence Nationale de la Recherche ([ANR-18-IAHU-0001)

  • Filippo Del Bene

Fondation pour la Recherche Médicale (ECO20170637481)

  • Marion Rosello

Ligue Nationale Contre le Cancer

  • Marion Rosello

UNADEV/AVIESAN

  • Filippo Del Bene

World Wide Cancer Research (grant no. 0624)

  • Marina C Mione

LILT -Trento (Program 5 per mille)

  • Marina C Mione

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

Ethics

Animal experimentation: All procedures were performed on zebrafish embryos in accordance with the European Communities Council Directive (2010/63/EU) and French law (87/848) and approved by the Sorbonne Université ethic committee (Charles Darwin) and the French Ministry for research (APAFIS agreement #21323 2019062416186982) and by the Institut Curie ethic committee and the French Ministry for research (APAFIS agreement #6031 2016070822342309)

Reviewing Editor

  1. Tanya T Whitfield, University of Sheffield, United Kingdom

Publication history

  1. Received: December 7, 2020
  2. Accepted: February 10, 2021
  3. Accepted Manuscript published: February 12, 2021 (version 1)
  4. Version of Record published: March 4, 2021 (version 2)

Copyright

© 2021, Rosello 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. Marion Rosello
  2. Juliette Vougny
  3. François Czarny
  4. Marina C Mione
  5. Jean-Paul Concordet
  6. Shahad Albadri
  7. Filippo Del Bene
(2021)
Precise base editing for the in vivo study of developmental signaling and human pathologies in zebrafish
eLife 10:e65552.
https://doi.org/10.7554/eLife.65552

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