基础医学与临床2013，Vol.33Issue(8)：986-991,6.
HCV5'UTR靶向性人工M1GS核酶的胞内抗病毒活性
The intracellular antiviral activities of a novel engineered M1GS ribozyme that targets to 5'untranslated region of hepatitis C virus genome
摘要
Abstract
Objective To explore new antiviral agents and therapies for hepatitis C.Methods The sequence and structure of the 5' untranslated region of HCV genome were analyzed with two computer software,DNAMAN and RNA Structure.The cytosine 67 nt downstream of the first base of HCV genome RNA was identified as the optimal target cleavage site.Based on the flanking sequence of this assumed cleavage site,a guide sequence (GS) was designed and covalently linked to the 3 prime terminus of the M1 RNA,which is a catalytic subunit of the RNase P derived from Escherichia coli using PCR.Results In the in vitro cleavage assay,The M1GS-HCV/C67 ribozyme could effectively cleave the HCV target RNA into two fragments at the specific cleavage site.Moreover,comparing to the blank control,this engineered M1 GS ribozyme could reduce the core protein expression of more than 75％ in the HCV-infected host cell and lead to a 800-fold reduction of HCV RNA copies in the culture supernatant.An another M1 GS ribozyme,M1 GS-HCV/C67 *,which has the same guide sequence but does not contain the bridge sequence,did not exhibit apparent inhibition for the expression of HCV core gene and viral proliferation in our paralleled assay.Conclusions The engineered ribozyme has notably antiviral activity in cultured cells,thus could provide a new promising approach for development of anti-HCV strategy.关键词
M1GS核酶/丙型肝炎病毒/5'非翻译区/抗病毒Key words
M1GS ribozyme/ HCV /5'UTR / antiviral分类
医药卫生引用本文复制引用
张文军,黄志文,李喜芳,张成成,刘映乐..HCV5'UTR靶向性人工M1GS核酶的胞内抗病毒活性[J].基础医学与临床,2013,33(8):986-991,6.基金项目
广东省自然科学基金(S2012010009471) （S2012010009471）
病毒学国家重点实验室开放基金(2012008) （2012008）
