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Highly efficient gene knockout in mice and zebrafish with RNA-guided endonucleases
Cited 219 time in
Web of Science
Cited 236 time in Scopus
- Authors
- Issue Date
- 2014-01
- Publisher
- Cold Spring Harbor Laboratory Press
- Citation
- Genome Research, Vol.24 No.1, pp.125-131
- Abstract
- RNA-guided endonucleases (RGENs), derived from the prokaryotic Type II CRISPR-Cas system, enable targeted genome modification in cells and organisms. Here we describe the establishment of gene-knockout mice and zebrafish by the injection of RGENs as Cas9 protein: guide RNA complexes or Cas9 mRNA plus guide RNA into one-cell-stage embryos of both species. RGENs efficiently generated germline transmittable mutations in up to 93% of newborn mice with minimal toxicity. RGEN-induced mutations in the mouse Prkdc gene that encodes an enzyme critical for DNA double-strand break repair resulted in immunodeficiency both in F-0 and F-1 mice. We propose that RGEN-mediated mutagenesis in animals will greatly expedite the creation of genetically engineered model organisms, accelerating functional genomic research.
- ISSN
- 1088-9051
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