PUBLICATION
Precise and efficient genome editing in zebrafish using the CRISPR/Cas9 system
- Authors
- Irion, U., Krauss, J., Nüsslein-Volhard, C.
- ID
- ZDB-PUB-141121-1
- Date
- 2014
- Source
- Development (Cambridge, England) 141(24): 4827-30 (Journal)
- Registered Authors
- Irion, Uwe, Krauss, Jana, Nüsslein-Volhard, Christiane
- Keywords
- none
- MeSH Terms
-
- Zebrafish/genetics*
- Genetic Engineering/methods*
- DNA Primers/genetics
- Codon, Nonsense/genetics*
- Polymorphism, Single Nucleotide/genetics
- Zebrafish Proteins/genetics*
- DNA Repair/genetics*
- Clustered Regularly Interspaced Short Palindromic Repeats/genetics
- Polymerase Chain Reaction
- Animals
- DNA, Circular/genetics
- Genotype
- Genome/genetics*
- Membrane Transport Proteins/genetics*
- CRISPR-Associated Proteins/genetics
- PubMed
- 25411213 Full text @ Development
Citation
Irion, U., Krauss, J., Nüsslein-Volhard, C. (2014) Precise and efficient genome editing in zebrafish using the CRISPR/Cas9 system. Development (Cambridge, England). 141(24):4827-30.
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping