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

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PC4 RNAi靶序列。

Abstract: 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-293T cell 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.