Establishment of Ace2 knockout mouse model with CRISPR/Cas9 gene targeting technology / 生理学报
Acta Physiologica Sinica
;
(6): 588-596, 2019.
Article
in Chinese
| WPRIM
| ID: wpr-777152
ABSTRACT
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 mice of F3 generation. Ace2 mRNA and protein were not detected in tissues of these Ace2 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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Mice, Knockout
/
Gene Targeting
/
Gene Knockout Techniques
/
CRISPR-Cas Systems
/
Genetics
Limits:
Animals
Language:
Chinese
Journal:
Acta Physiologica Sinica
Year:
2019
Type:
Article
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