title: Creation of mutant mice with megabase-sized deletions containing custom-designed breakpoints by means of the CRISPR/Cas9 system.
abstract: The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system is a useful tool for creation of mutant mice with mutations mirroring those in human patients. Various methods have been developed for this purpose, including deletions, inversions, and translocations. So far, mutant mice with deletions of up to 1.2 megabases (Mb) have been generated by microinjection of the CRISPR/Cas9 system into fertilized eggs; however, a method for generation of mutant mice with a deletion of more than several Mb size is necessary because such deletions have often been identified as possible causes of human diseases. With an aim to enable the generation of disease models carrying large deletions with a breakpoint in custom-designed sequences, we developed a method for induction of an Mb-sized deletion by microinjection of a pair of sgRNAs, Cas9, and a donor plasmid into fertilized eggs. Using this method, we efficiently and rapidly generated mutant mice carrying deletions up to 5 Mb.
authors.affiliation: Department of Systems BioMedicine, National Research Institute for Child Health and Development, Tokyo, 157-8535, Japan.
authors.forename: Tomoko
authors.initials: T
authors.lastname: Kato
authors.affiliation: Department of Systems BioMedicine, National Research Institute for Child Health and Development, Tokyo, 157-8535, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Hara
authors.affiliation: Department of Biology, Faculty of Science, Toho University, Miyama 2-2-1, Funabashi, Chiba, 274-8510, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Goto
authors.affiliation: Department of Systems BioMedicine, National Research Institute for Child Health and Development, Tokyo, 157-8535, Japan.|Department of Biology, Faculty of Science, Toho University, Miyama 2-2-1, Funabashi, Chiba, 274-8510, Japan.
authors.forename: Yuya
authors.initials: Y
authors.lastname: Ogawa
authors.affiliation: Department of Systems BioMedicine, National Research Institute for Child Health and Development, Tokyo, 157-8535, Japan.
authors.forename: Haruka
authors.initials: H
authors.lastname: Okayasu
authors.affiliation: Department of Biology, Faculty of Science, Toho University, Miyama 2-2-1, Funabashi, Chiba, 274-8510, Japan.
authors.forename: Souichirou
authors.initials: S
authors.lastname: Kubota
authors.affiliation: Department of Systems BioMedicine, National Research Institute for Child Health and Development, Tokyo, 157-8535, Japan.
authors.forename: Moe
authors.initials: M
authors.lastname: Tamano
authors.affiliation: Department of Systems BioMedicine, National Research Institute for Child Health and Development, Tokyo, 157-8535, Japan.
authors.forename: Miho
authors.initials: M
authors.lastname: Terao
authors.affiliation: Department of Systems BioMedicine, National Research Institute for Child Health and Development, Tokyo, 157-8535, Japan. takada-s@ncchd.go.jp.
authors.forename: Shuji
authors.initials: S
authors.lastname: Takada
mesh_terms: D000818:Animals; D064113:CRISPR-Cas Systems; D004195:Disease Models, Animal; D005260:Female; D017353:Gene Deletion; D008297:Male; D008817:Mice, Mutant Strains / Q000235:genetics*; D008845:Microinjections; D009154:Mutation; D010957:Plasmids; D015053:Zygote