| First Author | Aida T | Year | 2015 |
| Journal | Genome Biol | Volume | 16 |
| Issue | 1 | Pages | 87 |
| PubMed ID | 25924609 | Mgi Jnum | J:345651 |
| Mgi Id | MGI:6316807 | Doi | 10.1186/s13059-015-0653-x |
| Citation | Aida T, et al. (2015) Cloning-free CRISPR/Cas system facilitates functional cassette knock-in in mice. Genome Biol 16:87 |
| abstractText | Although the CRISPR/Cas system has enabled one-step generation of knockout mice, low success rates of cassette knock-in limit its application range. Here we show that cloning-free, direct nuclear delivery of Cas9 protein complex with chemically synthesized dual RNAs enables highly efficient target digestion, leading to generation of knock-in mice carrying a functional cassette with up to 50% efficiency, compared with just 10% by a commonly used method consisting of Cas9 mRNA and single guide RNA. Our cloning-free CRISPR/Cas system facilitates rapid one-step generation of cassette knock-in mice, accelerating functional genomic research by providing various in vivo genetic tools. |
