| First Author | Liu C | Year | 2019 |
| Journal | Sheng Li Xue Bao | Volume | 71 |
| Issue | 4 | Pages | 588-596 |
| PubMed ID | 31440756 | Mgi Jnum | J:285491 |
| Mgi Id | MGI:6400383 | Citation | Liu C, et al. (2019) [Establishment of Ace2 knockout mouse model with CRISPR/Cas9 gene targeting technology]. Sheng Li Xue Bao 71(4):588-596 |
| abstractText | The aim of the study was to establish Ace2 (angiotensin-converting enzyme 2) knockout mouse model with CRISPR/Cas9 gene targeting technology. A vector targeting Ace2 gene knockout was constructed with the primers of single-guide RNA (gRNA), and then transcribed gRNA/Cas9 mRNA was micro-injected into the mouse zygote. The deletion of exons 3 to 18 of Ace2 gene in mice was detected and identified by PCR and gene sequencing. The Ace2 gene knock-out mice were bred and copulated. Ace2 protein and mRNA expression were detected by Western blot and qRT-PCR in F3 progeny knock-out male mice. The gRNA expression vector was successfully constructed and transcribed in vitro, and active gRNA and Cas9 mRNA were injected directly into zygote. The deletion of exons 3 to 18 of Ace2 gene in six positive founder mice as the F0 generation were confirmed by PCR and gene sequencing. Six founder mice were mated with wild-type mice, then achieved F1 generation were mated and produced F2 generation. The female positive mouse of F2 was selected to mate with wild-type mice and produce Ace2(-/Y) mice of F3 generation. Ace2 mRNA and protein were not detected in tissues of these Ace2(-/Y) mice. In conclusion, a mouse model with Ace2 deficiency has been successfully established with CRISPR/Cas9 technique, which shall lay a foundation for future investigation of Ace2. |
