RRC ID 49490
Author Kato T, Hara S, Goto Y, Ogawa Y, Okayasu H, Kubota S, Tamano M, Terao M, Takada S.
Title Creation of mutant mice with megabase-sized deletions containing custom-designed breakpoints by means of the CRISPR/Cas9 system.
Journal 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.
Volume 7(1)
Pages 59
Published 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
Times Cited 5
DNA material B6N Mouse BAC clone (RDB07573): B6Ng01-234K06 B6Ng01-223B06 B6Ng01-111E12 B6Ng01-299I16 B6Ng01-295D01 B6Ng01-223B06 B6Ng01-332K06 B6Ng01-111E12.