一个能够有效介导RNA干扰或外源基因过表达的慢病毒系统

A Lentiviral System Capable of Mediating RNA Interference or Overexpression of Foreign Genes

[目的]为研究pRRL为基础的慢病毒系统除了作为RNA干扰工具外是否可用于介导外源基因过表达。[方法]构建能表达NALP2蛋白N末端PYRIN结构域(PYD)或p53的慢病毒;采用RT-PCR、Western印迹法检测慢病毒介导的PYD、p53表达情况;采用NF-κB荧光素酶报告实验和细胞凋亡实验鉴定PYD和p53的生物学功能。[结果]通过对绿色荧光蛋白(EGFP)的观察和对PYD、p53表达水平的检测,表明成功构建了能够表达PYD或p53的慢病毒。NF-κB荧光素酶报告实验和细胞凋亡实验结果表明慢病毒表达的PYD和p53具有生物学功能。[结论]pRRL为基础的慢病毒系统除了能够作为RNA干扰工具外,还可介导外源基因的表达,并在体外高效感染肿瘤细胞。

[Purpose]To investigate whether the pRRL-based lentiviral system is capable of medi- ating overexpression of foreign genes besides RNA interference.[Methods]A lentiviral system expressing PYD or p53 was packaged.PYD or p53 expression was detected by RT-PCR and West- ern blot.The biological function of PYD and p53 was tested by NF-κB luciferase reporter assay and apoptosis assay respectively.[Results]By means of EGFP by determined and detecting levels of PYD or p53 expression,lentivirus expressing PYD or p53...