PC4基因RNAi慢病毒载体的构建及有效靶序列鉴定  

Construction and screening of effective sequences of lentiviral-mediated RNAi targeting PC4

作　　者：张真中[1] 杨春杰[1] 钱三立 李冰玉[1] 邹云增[1] ZHANG Zhen-zhong;YANG Chun-jie;QIAN San-li;LI Bing-yu;ZOU Yun-zeng(Shanghai Institute of Cardiovascular Diseases,Zhongshan Hospital,Fudan University,Shanghai 200032,China)

机构地区：[1]复旦大学附属中山医院上海市心血管病研究所,上海200032

出　　处：《中国临床医学》2018年第3期427-430,共4页Chinese Journal of Clinical Medicine

基　　金：国家自然基金重大国际(地区)合作研究项目(81220108003)~~

摘　　要：目的:筛选慢病毒介导PC4 RNA干扰的有效靶序列。方法:通过数据库设计3条PC4干扰候选靶序列,将靶序列构建到慢病毒载体中,转入HEK-293T细胞后,感染H9C2心肌细胞。通过实时荧光定量PCR和蛋白免疫印迹法鉴定靶序列干扰PC4的效果。结果:成功构建shPC4慢病毒载体,获得高效感染心肌细胞的慢病毒;感染心肌细胞后实时荧光定量PCR和蛋白免疫印迹显示,shPC4-1具有较高的干扰效果(P<0.05)。结论:成功筛选出高效的PC4RNAi靶序列。Objective：To screen the effective sequences of lentivirus-mediated RNAi targeting PC4.Methods：Three candidate target sequences of PC4 RNAi were designed and cloned into lentivirus vectors,packaged into HEK-293 Tcell line,and then transfected the H9C2 cell line.The interference effects of target sequences were evaluated by real-time PCR and Western blotting analysis.Results：The lentiviral vector of RNAi containing the sequences targeting PC4 were successfully constructed,and effectively cloned into the H9C2 cell line.Real-time PCR and Western blotting results showed that PC4 expression in H9C2 cell line was downregulated significantly by shPC4-1.Conclusions：Sequences of lentivirus-mediated RNAi of PC4 are successfully screened.

关 键 词：PC4 慢病毒 RNA干扰 心肌细胞 

分 类 号：R393[医药卫生—基础医学]