title: One-step generation of phenotype-expressing triple-knockout mice with heritable mutated alleles by the CRISPR/Cas9 system.
abstract: 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.
authors.affiliation: Department of Animal Resource Sciences, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan.
authors.forename: Wataru
authors.initials: W
authors.lastname: Fujii
authors.forename: Asuka
authors.initials: A
authors.lastname: Onuma
authors.forename: Koji
authors.initials: K
authors.lastname: Sugiura
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Naito
mesh_terms: D000483:Alleles; D000818:Animals; D064112:Clustered Regularly Interspaced Short Palindromic Repeats; D005260:Female; D055786:Gene Knockout Techniques; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009154:Mutation; D010641:Phenotype
references.citation: Nature. 1997 May 1;387(6628):83-90
references.pmid: 9139826
references.citation: Nat Biotechnol. 2013 Aug;31(8):684-6
references.pmid: 23929337
references.citation: PLoS One. 2007 Jun 27;2(6):e579
references.pmid: 17593978
references.citation: Biochem Biophys Res Commun. 2014 Mar 21;445(4):791-4
references.pmid: 24491566
references.citation: Cell. 2013 May 9;153(4):910-8
references.pmid: 23643243
references.citation: Genesis. 2005 Oct;43(2):71-9
references.pmid: 16100711
references.citation: Genomics. 1996 Oct 15;37(2):245-8
references.pmid: 8921397
references.citation: Int J Biochem Cell Biol. 2014 Jan;46:49-55
references.pmid: 24269190
references.citation: Nat Genet. 1993 Mar;3(3):235-40
references.pmid: 8485579
references.citation: Nucleic Acids Res. 2013 Nov;41(20):e187
references.pmid: 23997119
references.citation: Science. 2013 Feb 15;339(6121):823-6
references.pmid: 23287722
references.citation: Science. 2013 Feb 15;339(6121):819-23
references.pmid: 23287718