Reference - Detail
| RRC ID | 50502 |
|---|---|
| Author | Miyasaka Y, Uno Y, Yoshimi K, Kunihiro Y, Yoshimura T, Tanaka T, Ishikubo H, Hiraoka Y, Takemoto N, Tanaka T, Ooguchi Y, Skehel P, Aida T, Takeda J, Mashimo T. |
| Title | CLICK: one-step generation of conditional knockout mice. |
| Journal | BMC Genomics |
| Abstract |
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. |
| Volume | 19(1) |
| Pages | 318 |
| Published | 2018-5-2 |
| DOI | 10.1186/s12864-018-4713-y |
| PII | 10.1186/s12864-018-4713-y |
| PMID | 29720086 |
| PMC | PMC5930688 |
| MeSH | Alleles Animals Base Sequence CRISPR-Cas Systems / genetics Genetic Engineering / methods* Injections Mice Mice, Knockout Zygote / metabolism |
| IF | 3.594 |
| Resource | |
| Mice | RBRC01828 |