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目的:构建p53凋亡刺激蛋白2(ASPP2)基因敲除小鼠,用该小鼠建立二乙基亚硝胺(DEN)诱导的肝癌模型以研究ASPP2的生物学功能。方法:构建单链向导RNA寡聚核苷酸,用CRISPR/Cas9系统制备ASPP2基因敲除小鼠。采用PCR法和测序方法鉴定F0、F1代及其子代的基因型。用DEN诱导ASPP2+/-小鼠建立肝癌模型。结果:PCR和测序结果表明F0代小鼠ASPP2基因敲除成功;F1代小鼠基因型符合ASPP2+/-,获得稳定遗传性;F1代自杂交获得ASPP2+/-小鼠DEN诱导的肝癌模型成功率(7/8与3/8)明显高于野生型。结论:基于CRISPR/Cas9系统成功构建ASPP2基因敲除小鼠,ASPP2基因敲除小鼠DEN诱导的肝癌模型成功率明显高于野生型。“,”Objective: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.Methods: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.Results: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.Conclusion: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.