RRC ID 39659
Author Hara S, Kato T, Goto Y, Kubota S, Tamano M, Terao M, Takada S.
Title Microinjection-based generation of mutant mice with a double mutation and a 0.5 Mb deletion in their genome by the CRISPR/Cas9 system.
Journal 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.
Volume 62(5)
Pages 531-536
Published 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
Times Cited 9
WOS Category AGRICULTURE, DAIRY & ANIMAL SCIENCE REPRODUCTIVE BIOLOGY
Resource
DNA material B6N BAC Mouse (RDB07573) B6Ng01-076D03 B6Ng01-096G05 B6Ng01-008H07 B6Ng01-289E22.