SiRNA干扰HSV-1即刻早期基因α22效应的初步分析

Analysis of siRNA Interfering in the α22 Immediate-early Gene of HSV-1

以HSV1即刻早期基因α22为靶点,应用T7RNA聚合酶体外合成siRNA1和siRNA2序列,脂质体转染法将其导入Hep2或KMB17细胞,再接种HSV1病毒,通过细胞形态和病毒滴度的测定,初步探索了siRNA干扰α22基因的的效应。结果表明,在Hep2中,转染siRNA的实验组与对照组的细胞病变时间相同,子代病毒的生长曲线也没有显著差异;而分别转染或共同转染siRNA1和siRNA2于“限制"性细胞KMB17中,接种病毒后,ICP22特异的siRNA对HSV1的复制则表现了相应的干扰作用,细胞病变慢,子代病毒的最高滴度峰值出现较晚。

In this study, α22, an immediate-early gene of HSV-1 was analyzed with RNAi technology. Two sequences of siRNAs (siRNA-1 and siRNA-2) were prepared by in vitro transcription, and transfected into the Hep-2 and KMB17 cells. The cells were subsequently inoculated with HSV-1. The results demonstrated that there was no differences between the transfected Hep-2 and the untransfected cells. Meanwhile, KMB-17, a kind of restricted cell, which was transfected respectively or co-transfected with siRNA-1 and siRNA-2, showed delayed cytopathic effects with the peak titre of offspring virus appearing later. The results suggested the two siRNAs can interfere with the expression of ICP22 and further effect on the replication of HSV-1.