Inhibition of hepatitis C virus replication by pol III-directed overexpression of RNA decoys corresponding to stem-loop structures in the NS5B coding region

Jing Zhang, Osamu Yamada, Takashi Sakamoto, Hiroshi Yoshida, Hiromasa Araki, Takayuki Murata, Kunitada Shimotohno

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

Increasing evidence has shown that the stem-loop (SL) structures in the NS5B coding region of hepatitis C virus (HCV) function as cis-replicating elements that are indispensable for viral replication. We have investigated whether small RNA molecules analogous to the SL structures could inhibit HCV replication. Reporter assays showed that both in vitro transcribed and pol III-directed transcripts corresponding to 5BSL3.1 and 5BSL3.2 efficiently inhibited HCV replicon-encoded luciferase expression. Mutagenesis studies revealed that mutation in 5BSL3.2 which debilitated its binding to NS5B also abolished the ability of 5BSL3.2 RNA to inhibit HCV replication, suggesting that SL RNA inhibits HCV by sequestering the replication complex. Further, adenoviral-mediated expression of the SL RNAs potently blocked the replication of HCV replicon in Huh-7 cells. Importantly, SL RNAs derived from HCV 2a, an evolutionarily distant genotype, were also shown to suppress the replication of HCV 1b replicon in spite of the genetic heterogeneity between the SL elements of the two viruses, implying the potential of SL RNA-based approach to inhibit a wide range of HCV isolates. These results suggest that SL RNA decoys may prove to be useful in the treatment of hepatitis C, which may be advantageous over other sequence-specific gene therapy modalities (such as antisense RNA and siRNA) in preventing the escape of genetic variants.

Original languageEnglish
Pages (from-to)276-285
Number of pages10
JournalVirology
Volume342
Issue number2
DOIs
Publication statusPublished - 25-11-2005
Externally publishedYes

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Virus Replication
Hepacivirus
RNA
Replicon
Antisense RNA
Genetic Heterogeneity
Hepatitis C
Luciferases
Mutagenesis
Genetic Therapy
Small Interfering RNA
Genotype
Viruses
Mutation

All Science Journal Classification (ASJC) codes

  • Virology

Cite this

Zhang, Jing ; Yamada, Osamu ; Sakamoto, Takashi ; Yoshida, Hiroshi ; Araki, Hiromasa ; Murata, Takayuki ; Shimotohno, Kunitada. / Inhibition of hepatitis C virus replication by pol III-directed overexpression of RNA decoys corresponding to stem-loop structures in the NS5B coding region. In: Virology. 2005 ; Vol. 342, No. 2. pp. 276-285.
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Inhibition of hepatitis C virus replication by pol III-directed overexpression of RNA decoys corresponding to stem-loop structures in the NS5B coding region. / Zhang, Jing; Yamada, Osamu; Sakamoto, Takashi; Yoshida, Hiroshi; Araki, Hiromasa; Murata, Takayuki; Shimotohno, Kunitada.

In: Virology, Vol. 342, No. 2, 25.11.2005, p. 276-285.

Research output: Contribution to journalArticle

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