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Journal Article

Precise and efficient genome editing in zebrafish using the CRISPR/Cas9 system

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Irion,  U
Department Genetics, Max Planck Institute for Developmental Biology, Max Planck Society;

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Krauss,  J
Department Genetics, Max Planck Institute for Developmental Biology, Max Planck Society;

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Nüsslein-Volhard,  C
Department Genetics, Max Planck Institute for Developmental Biology, Max Planck Society;

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Citation

Irion, U., Krauss, J., & Nüsslein-Volhard, C. (2014). Precise and efficient genome editing in zebrafish using the CRISPR/Cas9 system. Development, 141(24), 4827-4830. doi:10.1242/dev.115584.


Cite as: https://hdl.handle.net/21.11116/0000-000A-7E64-9
Abstract
The introduction of engineered site-specific DNA endonucleases has brought precise genome editing in many model organisms and human cells into the realm of possibility. In zebrafish, loss-of-function alleles have been successfully produced; however, germ line transmission of functional targeted knock-ins of protein tags or of SNP exchanges have not been reported. Here we show by phenotypic rescue that the CRISPR/Cas system can be used to target and repair a premature stop codon at the albino (alb) locus in zebrafish with high efficiency and precision. Using circular donor DNA containing CRISPR target sites we obtain close to 50% of larvae with precise homology-directed repair of the alb(b4) mutation, a small fraction of which transmitted the repaired allele in the germ line to the next generation (3/28 adult fish). The in vivo demonstration of germ line transmission of a precise SNP exchange in zebrafish underscores its suitability as a model for genetic research.