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Originally published as JGV in Press, 10.1099/vir.0.011338-0 on June 24, 2009 J Gen Virol 90 (2009), 2468-2473; DOI 10.1099/vir.0.011338-0

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Short Communication

Inhibition of coxsackievirus B3 and related enteroviruses by antiviral short interfering RNA pools produced using {phi}6 RNA-dependent RNA polymerase

Michaela Nygårdas1, Tytti Vuorinen1, Antti P. Aalto2, Dennis H. Bamford2 and Veijo Hukkanen1,3

1 Department of Virology, University of Turku, Kiinamyllynkatu 13, FIN-20520 Turku, Finland
2 Institute of Biotechnology and Department of Biological and Environmental Sciences, Biocenter 2, Viikinkaari 5, PO Box 56, FIN-00014 University of Helsinki, Finland
3 Department of Microbiology, Aapistie 5A, 90014 University of Oulu, Finland

Correspondence
Michaela Nygårdas
mnygarda{at}abo.fi

Coxsackievirus B3 (CBV3) is a member of the human enterovirus B species and a common human pathogen. Even though much is known about the enteroviral life cycle, no specific drugs are available to treat enterovirus infections. RNA interference (RNAi) has evolved to be an important tool for antiviral experimental therapies and gene function studies. We describe here a novel approach for RNAi against CBVs by using a short interfering (siRNA) pool covering 3.5 kb of CBV3 genomic sequence. The RNA-dependent RNA polymerase (RdRP) of bacteriophage {phi}6 was used to synthesize long double-stranded RNA (dsRNA) from a cloned region (nt 3837–7399) of the CBV3 genome. The dsRNA was cleaved using Dicer, purified and introduced to cells by transfection. The siRNA pool synthesized using the {phi}6 RdRP ({phi}6–siRNAs) was considerably more effective than single-site siRNAs. The {phi}6–siRNA pool also inhibited replication of other enterovirus B species, such as coxsackievirus B4 and coxsackievirus A9.

A supplementary table is available with the online version of this paper.






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