CRISPR pp 317-334 | Cite as

Targeted Mutagenesis in Zebrafish Using CRISPR RNA-Guided Nucleases

  • Woong Y. Hwang
  • Yanfang Fu
  • Deepak Reyon
  • Andrew P. W. Gonzales
  • J. Keith Joung
  • Jing-Ruey Joanna Yeh
Part of the Methods in Molecular Biology book series (MIMB, volume 1311)

Abstract

In recent years, the zebrafish has become a critical contributor to various areas of biomedical research, advancing our fundamental understanding of biomedicine and helping discover candidate therapeutics for human diseases. Nevertheless, to further extend the power of this important model organism requires a robust and simple-to-use genome editing platform that will enable targeted gene knockouts and introduction of specific mutations identified in human diseases into the zebrafish genome. We describe here protocols for creating insertion or deletion (indel) mutations or precise sequence modifications in zebrafish genes using customizable CRISPR-Cas9 RNA-guided nucleases (RGNs). These methods can be easily implemented in any lab and may also potentially be extended for use in other organisms.

Key words

CRISPR Cas9 Gene-editing Genome engineering Zebrafish T7E1 

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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Woong Y. Hwang
    • 1
    • 2
  • Yanfang Fu
    • 3
    • 4
    • 5
  • Deepak Reyon
    • 3
    • 4
    • 5
  • Andrew P. W. Gonzales
    • 1
    • 2
  • J. Keith Joung
    • 3
    • 4
    • 5
  • Jing-Ruey Joanna Yeh
    • 1
    • 2
  1. 1.Cardiovascular Research CenterMassachusetts General HospitalCharlestownUSA
  2. 2.Department of MedicineHarvard Medical SchoolBostonUSA
  3. 3.Molecular Pathology Unit, Center for Cancer ResearchMassachusetts General HospitalCharlestownUSA
  4. 4.Center for Computational and Integrative BiologyMassachusetts General HospitalCharlestownUSA
  5. 5.Department of PathologyHarvard Medical SchoolBostonUSA

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