RRC ID |
39659
|
著者 |
Hara S, Kato T, Goto Y, Kubota S, Tamano M, Terao M, Takada S.
|
タイトル |
Microinjection-based generation of mutant mice with a double mutation and a 0.5 Mb deletion in their genome by the CRISPR/Cas9 system.
|
ジャーナル |
J Reprod Dev
|
Abstract |
The clustered regularly interspaced short palindromic repeat (CRISPR)/CRISPR-associated protein 9 (Cas9) system is a useful tool for genome editing. In this study, using a microinjection-based CRISPR/Cas9 system, we efficiently generated mouse lines carrying mutations at the Irx3 and Irx5 loci, which are located in close proximity on a chromosome and are functionally redundant. During the generation of Irx3/Irx5 double mutant mice, a deletion of ~0.5 Mb between the Irx3 and Irx5 loci was unintentionally identified in 6 out of 27 living pups by PCR based genotyping analysis. This deletion was confirmed by DNA fluorescence in situ hybridization analysis of fibroblasts. These results indicate that the mutant mice with a deletion of at least 0.5 Mb in their genome can be generated by the CRISPR/Cas9 system through microinjection into fertilized eggs. Our findings expand the utility of the CRISPR/Cas9 system in production of disease model animals with large deletions.
|
巻・号 |
62(5)
|
ページ |
531-536
|
公開日 |
2016-10-18
|
DOI |
10.1262/jrd.2016-058
|
PMID |
27396308
|
PMC |
PMC5081742
|
MeSH |
Alleles
Animals
CRISPR-Cas Systems*
Codon
Exons
Female
Fibroblasts / metabolism
Gene Deletion*
Genetic Vectors
Genome
Genotype
Homeodomain Proteins / genetics
In Situ Hybridization, Fluorescence
Mice
Mice, Inbred C57BL
Mice, Inbred DBA
Microinjections
Mutation*
Sequence Analysis, DNA
Transcription Factors / genetics
|
IF |
1.735
|
引用数 |
9
|
WOS 分野
|
AGRICULTURE, DAIRY & ANIMAL SCIENCE
REPRODUCTIVE BIOLOGY
|
リソース情報 |
遺伝子材料 |
B6N BAC Mouse (RDB07573) B6Ng01-076D03
B6Ng01-096G05
B6Ng01-008H07
B6Ng01-289E22. |