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


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.

SUBMITTER: Irion U 

PROVIDER: S-EPMC4299274 | biostudies-other | 2014 Dec

REPOSITORIES: biostudies-other

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

Irion Uwe U   Krauss Jana J   Nüsslein-Volhard Christiane C  

Development (Cambridge, England) 20141119 24


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 (al  ...[more]

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