Interference of hepatitis C virus replication in cell culture by antisense peptide nucleic acids targeting the X-RNA

被引:17
|
作者
Ahn, D. -G.
Shim, S. -B.
Moon, J. -E.
Kim, J. -H.
Kim, S. -J.
Oh, J. -W. [1 ]
机构
[1] Yonsei Univ, Dept Biotechnol, Seoul 120479, South Korea
基金
新加坡国家研究基金会;
关键词
antisense nucleic acids; HCV; NS5B; peptide nucleic acids; RdRp; X-RNA; 3' NONTRANSLATED REGION; VIRAL-RNA; BINDING-PROTEIN; CDNA-CLONE; IN-VIVO; POLYMERASE; SEQUENCES; GENOME; STRAND; SITE;
D O I
10.1111/j.1365-2893.2010.01416.x
中图分类号
R57 [消化系及腹部疾病];
学科分类号
摘要
The RNA-dependent RNA polymerase (RdRp) of hepatitis C virus (HCV) is the essential catalytic enzyme for viral genome replication. It initiates minus-strand RNA synthesis from a highly conserved 98-nt sequence, called the X-RNA, at the 3'-end of the plus-strand viral genome. In this study, we evaluated the antiviral effects of peptide nucleic acids (PNAs) targeting the X-RNA. Our in vitro RdRp assay results showed that PNAs targeting the three major stem-loop (SL) domains of X-RNA can inhibit RNA synthesis initiation. Delivery of X-RNA-targeted PNAs by fusing the PNAs to cell-penetrating peptides (CPPs) into HCV-replicating cells effectively suppressed HCV replication. Electrophoretic mobility shift assays revealed that the PNA targeting the SL3 region at the 5'-end of X-RNA dissociated the viral RdRp from the X-RNA. Furthermore, delivery of the SL3-targeted PNA into HCV-infected cells resulted in the suppression of HCV RNA replication without activation of interferon beta expression. Collectively, our results indicate that the HCV X-RNA can be effectively targeted by CPP-fused PNAs to block RNA-protein and/or RNA-RNA interactions essential for viral RNA replication and identify X-RNA SL3 as an RdRp binding site crucial for HCV replication. In addition, the ability to inhibit RNA synthesis initiation by targeting HCV X-RNA using antisense PNAs suggests their promising therapeutic potential against HCV infection.
引用
收藏
页码:E298 / E306
页数:9
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