Inhibition of coxsackievirus B3 replication by small interfering RNAs requires perfect sequence match in the central region of the viral positive strand

Ji Yuan, Paul K.M. Cheung, Huifang M. Zhang, David Chau, Decheng Yang

Research output: Contribution to journalArticle

81 Citations (Scopus)

Abstract

Coxsackievirus B3 (CVB3) is the most common causal agent of viral myocarditis, but existing drug therapies are of limited value. Application of small interfering RNA (siRNA) in knockdown of gene expression is an emerging technology in antiviral gene therapy. To investigate whether RNA interference (RNAi) can protect against CVB3 infection, we evaluated the effects of RNAi on viral replication in HeLa cells and murine cardiomyocytes by using five CVB3-specific siRNAs targeting distinct regions of the viral genome. The most effective one is siRNA-4, targeting the viral protease 2A, achieving a 92% inhibition of CVB3 replication. The specific RNAi effects could last at least 48 h, and cell viability assay revealed that 90% of siRNA-4-pretreated cells were still alive and lacked detectable viral protein expression 48 h postinfection. Moreover, administration of siRNAs after viral infection could also effectively inhibit viral replication, indicating its therapeutic potential. Further evaluation by combination found that no enhanced inhibitory effects were observed when siRNA-4 was cotransfected with each of the other four candidates. In mutational analysis of the mechanisms of siRNA action, we found that siRNA functions by targeting the positive strand of virus and requires a perfect sequence match in the central region of the target, but mismatches were more tolerated near the 3′ end than the 5′ end of the antisense strand. These findings reveal an effective target for CVB3 silencing and provide a new possibility for antiviral intervention.

Original languageEnglish (US)
Pages (from-to)2151-2159
Number of pages9
JournalJournal of virology
Volume79
Issue number4
DOIs
StatePublished - Feb 1 2005

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Enterovirus
small interfering RNA
Small Interfering RNA
RNA Interference
RNA interference
virus replication
Antiviral Agents
Coxsackievirus Infections
myocarditis
gene therapy
Viral Genome
viral proteins
Myocarditis
Viral Proteins
Virus Diseases
HeLa Cells
Cardiac Myocytes
infection
Genetic Therapy
drug therapy

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Inhibition of coxsackievirus B3 replication by small interfering RNAs requires perfect sequence match in the central region of the viral positive strand. / Yuan, Ji; Cheung, Paul K.M.; Zhang, Huifang M.; Chau, David; Yang, Decheng.

In: Journal of virology, Vol. 79, No. 4, 01.02.2005, p. 2151-2159.

Research output: Contribution to journalArticle

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