| RRC ID |
83364
|
| Author |
Yoshimi K, Kuno A, Yamauchi Y, Hattori K, Taniguchi H, Mikamo K, Iida R, Ishida S, Goto M, Takeshita K, Ito R, Takahashi R, Takahashi S, Mashimo T.
|
| Title |
Genome editing using type I-E CRISPR-Cas3 in mice and rat zygotes.
|
| Journal |
Cell Rep Methods
|
| Abstract |
The type I CRISPR system has recently emerged as a promising tool, especially for large-scale genomic modification, but its application to generate model animals by editing zygotes had not been established. In this study, we demonstrate genome editing in zygotes using the type I-E CRISPR-Cas3 system, which efficiently generates deletions of several thousand base pairs at targeted loci in mice with 40%-70% editing efficiency without off-target mutations. To overcome the difficulties associated with detecting the variable deletions, we used a newly long-read sequencing-based multiplex genotyping approach. Demonstrating remarkable versatility, our Cas3-based technique was successfully extended to rats as well as mice, even by zygote electroporation methods. Knockin for SNP exchange and genomic replacement with a donor plasmid were also achieved in mice. This pioneering work with the type I CRISPR zygote editing system offers increased flexibility and broader applications in genetic engineering across different species.
|
| Volume |
4(8)
|
| Pages |
100833
|
| Published |
2024-8-19
|
| DOI |
10.1016/j.crmeth.2024.100833
|
| PII |
S2667-2375(24)00206-6
|
| PMID |
39121862
|
| PMC |
PMC11384072
|
| MeSH |
Animals
CRISPR-Cas Systems* / genetics
Female
Gene Editing* / methods
Mice
Rats
Zygote* / metabolism
|
| Resource |
| Rats |
F344-Il2rgem1Larc (StrainID=1381)
F344-Rag2em1Larc (StrainID=1382) |
