特异性干扰ICP27基因的siRNA对HSV-1病毒复制的影响  

Inhibition of HSV-1 Virus Production by siRNAs Specific for ICP27 Gene

作　　者：王晓婷[1] 董慧[2] 王军[1] 全金梅[1] 张伟 

机构地区：[1]大连市友谊医院老年病科,中国辽宁大连116100 [2]大连市中心医院神经内科,中国辽宁大连116033 [3]大连医科大学基础医学院微生物学教研室,中国辽宁大连116044

出　　处：《生命科学研究》2017年第2期130-135,共6页Life Science Research

摘　　要：针对单纯疱疹病毒1型(herpes simplex virus 1,HSV-1)的ICP27基因和其他疱疹病毒相关基因的高度保守区设计小干扰RNA(small interfering RNA,siRNA),研究其抑制病毒复制的效果。首先构建相应的小发夹RNA(small hairpin RNA,shRNA),然后通过病毒滴度测定、real-time PCR和细胞致病变效应(cytopathic effect,CPE)检测所设计的siRNA抑制病毒复制的能力。结果显示,所设计的shRNA-2(靶序列起始位置815)和shRNA-3(靶序列起始位置1 367)具有明显地抑制病毒复制的效果。尤其是shRNA-3,抑制病毒复制的效果更明显,在病毒滴度实验中,与阴性对照相比,其抑制倍数为81,同时可以下调ICP27基因的mR NA表达水平。实验结果表明shRNA-3能够显著抑制HSV-1病毒复制的能力,可以作为HSV-1感染性疾病的补充治疗手段,其对应的靶序列可以作为抗HSV-1新的靶标。Small interfering RNAs （siRNAs） targeting ICP27 gene for herpes simplex virus 1 （HSV-1） and other highly conserved sequences for herpesviruses were designed and detected for their abilities to inhibit herpesvirus replication in vitro. Virus titer assay, real-time PCR and cytopathic effect （CPE） experiment were used to observe their inhibition effect in Vero cells challenged with HSV-1 by three small hairpin RNAs （shRNA-1, shRNA-2 and shRNA-3）. The results suggested shRNA-2 （target sequence from 815 POsition） and shRNA-3 （target sequence from 1 367 position） could depress the replication of HSV-1. Especially, virus repli- cation was obviously inhibited by shRNA-3 in the virus titer assay, and the inhibition ratio was 81 times comparing with Negative group. At the same time, the mRNA expression level of ICP27 gene could be decreased by shRNA-3. It indicated that shRNA-3 could effectively suppress the replication of HSV-1 and be used as a supplementary therapy for HSV-1 infectious diseases. The corresponding target sequence of shRNA-3 could therefore be used as a new target for anti-HSV-1 drugs.

关 键 词：单纯疱疹病毒1型(HSV-1) 小干扰RNA(siRNA) RNA干扰(RNAI) ICP27基因 

分 类 号：Q936[生物学—微生物学] R373.9[医药卫生—病原生物学]