| First Author | Aida T | Year | 2016 |
| Journal | BMC Genomics | Volume | 17 |
| Issue | 1 | Pages | 979 |
| PubMed ID | 27894274 | Mgi Jnum | J:239400 |
| Mgi Id | MGI:5828691 | Doi | 10.1186/s12864-016-3331-9 |
| Citation | Aida T, et al. (2016) Gene cassette knock-in in mammalian cells and zygotes by enhanced MMEJ. BMC Genomics 17(1):979 |
| abstractText | BACKGROUND: Although CRISPR/Cas enables one-step gene cassette knock-in, assembling targeting vectors containing long homology arms is a laborious process for high-throughput knock-in. We recently developed the CRISPR/Cas-based precise integration into the target chromosome (PITCh) system for a gene cassette knock-in without long homology arms mediated by microhomology-mediated end-joining. RESULTS: Here, we identified exonuclease 1 (Exo1) as an enhancer for PITCh in human cells. By combining the Exo1 and PITCh-directed donor vectors, we achieved convenient one-step knock-in of gene cassettes and floxed allele both in human cells and mouse zygotes. CONCLUSIONS: Our results provide a technical platform for high-throughput knock-in. |
