RRC ID 49490
著者 Kato T, Hara S, Goto Y, Ogawa Y, Okayasu H, Kubota S, Tamano M, Terao M, Takada S.
タイトル Creation of mutant mice with megabase-sized deletions containing custom-designed breakpoints by means of the CRISPR/Cas9 system.
ジャーナル Sci Rep
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.
巻・号 7(1)
ページ 59
公開日 2017-3-3
DOI 10.1038/s41598-017-00140-9
PII 10.1038/s41598-017-00140-9
PMID 28246396
PMC PMC5427885
MeSH Animals CRISPR-Cas Systems* Disease Models, Animal Female Gene Deletion* Male Mice, Mutant Strains / genetics* Microinjections Mutation* Plasmids Zygote
IF 3.998
引用数 5
リソース情報
遺伝子材料 B6N Mouse BAC clone (RDB07573): B6Ng01-234K06 B6Ng01-223B06 B6Ng01-111E12 B6Ng01-299I16 B6Ng01-295D01 B6Ng01-223B06 B6Ng01-332K06 B6Ng01-111E12.