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。 |
关键词: PC4 慢病毒 RNA干扰 心肌细胞 |
DOI: |
分类号:R 393 |
基金项目:国家自然基金重大国际(地区)合作研究项目(81220108003) |
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Construction and screening of effective sequences of lentiviral-mediated RNAi targeting PC4 |
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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. |
Key words: PC4 lentivirus RNA interference cardiomyocytes |