Highly efficient gene knockout in mice and zebrafish with RNA-guided endonucleases
- Authors
- Sung, Y.H., Kim, J.M., Kim, H.T., Lee, J., Jeon, J., Jin, Y., Choi, J.H., Ban, Y.H., Ha, S.J., Kim, C.H., Lee, H.W., and Kim, J.S.
- ID
- ZDB-PUB-140102-2
- Date
- 2014
- Source
- Genome research 24(1): 125-31 (Journal)
- Registered Authors
- Choi, Jung-Hwa
- Keywords
- none
- MeSH Terms
-
- Animals
- Animals, Newborn/genetics
- CRISPR-Associated Proteins/metabolism*
- Clustered Regularly Interspaced Short Palindromic Repeats
- Embryo, Mammalian
- Embryo, Nonmammalian
- Endonucleases/genetics*
- Endonucleases/metabolism
- Forkhead Transcription Factors/metabolism
- Gene Knockout Techniques
- Genome
- Germ-Line Mutation
- Mice
- Mice, Inbred BALB C
- Mice, Knockout
- Mutagenesis*
- Nuclear Proteins/genetics*
- Nuclear Proteins/metabolism
- Phenotype
- RNA, Guide, Kinetoplastida/genetics
- RNA, Guide, Kinetoplastida/metabolism
- Zebrafish/genetics
- Zebrafish/metabolism
- PubMed
- 24253447 Full text @ Genome Res.
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 F0 and F1 mice. We propose that RGEN-mediated mutagenesis in animals will greatly expedite the creation of genetically engineered model organisms, accelerating functional genomic research.