One-step generation of phenotype-expressing triple-knockout mice with heritable mutated alleles by the CRISPR/Cas9 system

J Reprod Dev. 2014;60(4):324-7. doi: 10.1262/jrd.2013-139. Epub 2014 May 4.


Triple-knockout mice generated by the one-step CRISPR/Cas9 system were examined for the effects of multiple gene modifications on each phenotype and individual gene function. Sixty embryos were transferred, and 9 pups were obtained; all 9 pups had mutations on 3 loci, and 7 pups showed mutations in all-alleles. F0 mice showed knockout phenotypes or no protein expression of target genes simultaneously, and these mutations were normally inherited in the next generation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Animals
  • Clustered Regularly Interspaced Short Palindromic Repeats
  • Female
  • Gene Knockout Techniques*
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mutation
  • Phenotype