| First Author | Miyasaka Y | Year | 2018 |
| Journal | BMC Genomics | Volume | 19 |
| Issue | 1 | Pages | 318 |
| PubMed ID | 29720086 | Mgi Jnum | J:261995 |
| Mgi Id | MGI:6156255 | Doi | 10.1186/s12864-018-4713-y |
| Citation | Miyasaka Y, et al. (2018) CLICK: one-step generation of conditional knockout mice. BMC Genomics 19(1):318 |
| abstractText | BACKGROUND: CRISPR/Cas9 enables the targeting of genes in zygotes; however, efficient approaches to create loxP-flanked (floxed) alleles remain elusive. RESULTS: Here, we show that the electroporation of Cas9, two gRNAs, and long single-stranded DNA (lssDNA) into zygotes, termed CLICK (CRISPR with lssDNA inducing conditional knockout alleles), enables the quick generation of floxed alleles in mice and rats. CONCLUSIONS: The high efficiency of CLICK provides homozygous knock-ins in oocytes carrying tissue-specific Cre, which allows the one-step generation of conditional knockouts in founder (F0) mice. |
