人大肠癌细胞系HCT116中Sirt1基因敲除细胞株的构建

摘要

目的 利用AAV介导的体细胞基因敲除技术,在人大肠癌细胞系HCT116中对靶基因Sirt 1进行敲除,达到基因沉默的目的.方法 构建Sirt1打靶载体,通过病毒包装、感染、药物筛选、PCR鉴定、Western blotting鉴定等步骤获得Sirt 1基因敲除的细胞株.结果 经过筛选和鉴定后获得两个Sirt1-/-阳性克隆,成功构建HCT116 Sirt1-/-细胞系.结论 通过体细胞敲除技术,我们成功地获得Sirt1敲除肠癌细胞株.%Objective To constructe of Sirt 1-/- cell line in HCT116 using the AAV knockout system. Methods We constructed targeting vector of Sirtl. After virus packaging, infecting, drug screening, PCR verification and western blotting, we got HCT116 Sirt-/-cell line. Results we constructed HCT116 Sirt 1-/- cell line successfully. Conclusion We successfully constructed the Sirtl - knockout cell line in HCT116.