人泡沫逆转录病毒介导的siRNA抑制HBV基因表达与复制  

Inhibition of Hepatitis B Virus by Small Interfering RNAs Expressed from Human Foamy Virus Vectors

作　　者：孙燕[1] 李利君[2] 刘辉[3] 李治[1] 李文鑫[3] 

机构地区：[1]陕西师范大学生命科学学院,陕西西安710062 [2]集美大学生物工程学院,福建厦门361021 [3]武汉大学生命科学学院/病毒学国家重点实验室,湖北武汉430072

出　　处：《武汉大学学报（理学版）》2007年第4期463-470,共8页Journal of Wuhan University:Natural Science Edition

基　　金：国家自然科学基金资助项目(30200083)

摘　　要：本研究采用基于PCR技术的siRNA表达方法,快速筛选到两个可以抑制HBV基因表达的siRNAs序列:S2和X1.S2和X1被克隆到人泡沫逆转录病毒(human foamy virus,HFV)载体中,构建成为分别表达S2和X1的单表达载体和同时表达S2和X1的双表达载体.将siRNAs表达载体转导入HepAD38细胞系中,多种检测分析的结果表明这些siRNA表达载体可以有效抑制多个HBV基因的表达和病毒DNA复制;并且其抑制作用是长期的,可以持续到转导后3个月.A polymerase chain reaction （PCR）-based siRNA expression strategy was employed to rapidly screen for effective siRNA sequences. Two effective siRNAs sequences （designated as S2 and X1） which reduced the HBV RNA by〉90% were identified. For delivering the siRNAs, they were cloned into a human foamy virus （HFV） based vector to generate single siRNA expression vectors HFVU6-siS2, HFVU6-siX1 and a dual siRNA expression vector HFVU6-siSX. After indrodcing the siRNAs into HepAD38 cell lines, the expression level of HBV genes and the replications of HBV were analyzed by ELISA, Western blot, Quantitative PCR analysis, RT-PCR and Southern blot. It was identified that these siRNA vectors can effectively inhibit multiple HBV gene expression and viral DNA replication. In addition, the repression of HBV RNA and DNA was stable up to 3 months posttransduction.

关 键 词：人泡沫逆转录病毒 病毒载体 RNA干扰 小干扰RNA 抗人乙肝病毒 

分 类 号：Q939.47[生物学—微生物学]