摘要
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.
作者
刘晓霓
寇卜心
柴梦音
陈德喜
Liu Xiaoni;Kou Buxin;Chai Mengyin;Chen Dexi(Beijing Institute of Hepatology,Beijing Youan Hospital,Capital Medical University,Beijing 100069,China)
出处
《中华肝脏病杂志》
CAS
CSCD
北大核心
2020年第9期784-789,共6页
Chinese Journal of Hepatology
基金
北京市自然科学基金(7192084)
北京市卫生系统高层次卫生技术人才培养计划(2015-3-101)
国家自然科学基金(81672026)。
