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Related Experiment Video

Updated: Jul 21, 2025

Efficient PAM-Less Base Editing for Zebrafish Modeling of Human Genetic Disease with zSpRY-ABE8e
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Precise mutagenesis in zebrafish using cytosine base editors.

Marion Rosello1, Malo Serafini2, Jean-Paul Concordet3

  • 1Sorbonne Université, INSERM U968, CNRS UMR 7210, Institut de la Vision, Paris, France. marion.rosello2@gmail.com.

Nature Protocols
|July 26, 2023
PubMed
Summary
This summary is machine-generated.

This study introduces efficient cytosine base editing in zebrafish, a powerful CRISPR tool for precise genome editing. This method accelerates the creation of zebrafish models for genetic research and disease studies.

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Area of Science:

  • Genetics
  • Molecular Biology
  • CRISPR Technology

Background:

  • CRISPR-based base editing enables precise genomic substitutions.
  • Generating precise point mutations in zebrafish remains a challenge.
  • Zebrafish are a key model for genetic and disease studies.

Purpose of the Study:

  • To describe experimental strategies and protocols for cytosine base editing in zebrafish.
  • To facilitate the generation of zebrafish lines with specific point mutations.
  • To optimize base editing efficiency and analyze phenotypic impacts.

Main Methods:

  • Guide RNA design and selection of base editor variants.
  • Utilizing alternative protospacer-adjacent motifs (PAMs) for broader targeting.
  • Employing co-selection via pigment gene mutations for efficient screening.
  • Direct phenotypic analysis of injected embryos.

Main Results:

  • Established protocols for cytosine base editing in zebrafish.
  • Demonstrated efficient generation of zebrafish mutants with desired substitutions.
  • Enabled rapid assessment of substitution effects within 2 weeks.

Conclusions:

  • Cytosine base editing is a powerful and efficient tool for zebrafish mutagenesis.
  • This method significantly advances in vivo genetic studies and disease modeling in zebrafish.
  • The described protocols streamline the generation and analysis of precise zebrafish mutants.