ZFIN ID: ZDB-PUB-141121-1
Precise and efficient genome editing in zebrafish using the CRISPR/Cas9 system
Irion, U., Krauss, J., Nüsslein-Volhard, C.
Date: 2014
Source: Development (Cambridge, England) 141(24): 4827-30 (Journal)
Registered Authors: Krauss, Jana, Nüsslein-Volhard, Christiane
Keywords: none
MeSH Terms:
  • Animals
  • CRISPR-Associated Proteins/genetics
  • Clustered Regularly Interspaced Short Palindromic Repeats/genetics
  • Codon, Nonsense/genetics*
  • DNA Primers/genetics
  • DNA Repair/genetics*
  • DNA, Circular/genetics
  • Genetic Engineering/methods*
  • Genome/genetics*
  • Genotype
  • Membrane Transport Proteins/genetics*
  • Polymerase Chain Reaction
  • Polymorphism, Single Nucleotide/genetics
  • Zebrafish/genetics*
  • Zebrafish Proteins/genetics*
PubMed: 25411213 Full text @ Development
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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.
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