RT Journal
A1 Sung, Young Hoon
A1 Kim, Jong Min
A1 Kim, Hyun-Taek
A1 Lee, Jaehoon
A1 Jeon, Jisun
A1 Jin, Young
A1 Choi, Jung-Hwa
A1 Ban, Young Ho
A1 Ha, Sang-Jun
A1 Kim, Cheol-Hee
A1 Lee, Han-Woong
A1 Kim, Jin-Soo
T1 Highly efficient gene knockout in mice and zebrafish with RNA-guided endonucleases
JF Genome Research 
JO Genome Research 
YR 2014 
FD January 01 
VO 24 
IS 1 
SP 125 
OP 131 
DO 10.1101/gr.163394.113 
UL http://genome.cshlp.org/content/24/1/125.abstract 
AB 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.