Liu X N, Kou B X, Chai M Y, Chen D X
Beijing Institute of Hepatology, Beijing Youan Hospital, Capital Medical University, Beijing 100069, China.
Zhonghua Gan Zang Bing Za Zhi. 2020 Sep 20;28(9):784-789. doi: 10.3760/cma.j.cn501113-20190728-00274.
To construct apoptosis-stimulating of p53 protein 2 (ASPP2) gene knockout mice using diethylnitrosamine (DEN)-induced liver cancer model to study the biological functions of ASPP2. The sgRNA oligonucleotides were constructed, and ASPP2 knockout mice were prepared with the CRISPR/Cas9 system. PCR and sequencing methods were used to identify the genotypes of F0 and F1 generations and their progeny. DEN was used to induce ASPP2+/- mice to establish liver cancer model. PCR and sequencing results showed that ASPP2 gene was successfully knocked out in F0 generation mice. The genotype of F1 generation mice was accorded with ASPP2+/- and had obtained stable heredity. The success rate of DEN-induced liver cancer model (7/8 and 3 / 8) of ASPP2 + /-mice obtained by self-hybridization of F1 generation was significantly higher than that of wild-type mice. ASPP2 knockout mice were successfully constructed based on the CRISPR/Cas9 system. The success rate of DEN-induced liver cancer model of ASPP2 knockout mice was significantly higher than that of the wild-type mice.
利用二乙基亚硝胺(DEN)诱导的肝癌模型构建p53蛋白凋亡刺激因子2(ASPP2)基因敲除小鼠,以研究ASPP2的生物学功能。构建sgRNA寡核苷酸,并用CRISPR/Cas9系统制备ASPP2基因敲除小鼠。采用PCR和测序方法鉴定F0和F1代及其后代的基因型。用DEN诱导ASPP2+/-小鼠建立肝癌模型。PCR和测序结果显示,F0代小鼠中ASPP2基因被成功敲除。F1代小鼠的基因型符合ASPP2+/-,并获得了稳定遗传。F1代自交获得的ASPP2+/-小鼠DEN诱导肝癌模型的成功率(7/8和3/8)显著高于野生型小鼠。基于CRISPR/Cas9系统成功构建了ASPP2基因敲除小鼠。ASPP2基因敲除小鼠DEN诱导肝癌模型的成功率显著高于野生型小鼠。