| 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
|
| Resource |
| DNA material |
B6N Mouse BAC clone (RDB07573): B6Ng01-234K06
B6Ng01-223B06
B6Ng01-111E12
B6Ng01-299I16
B6Ng01-295D01
B6Ng01-223B06
B6Ng01-332K06
B6Ng01-111E12. |
