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Silencing of African horse sickness virus VP7 protein expression in cultured cells by RNA interference

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Abstract

RNA interference (RNAi) is the process by which double-stranded RNA directs sequence-specific degradation of homologous mRNA. Short interfering RNAs (siRNAs) are the mediators of RNAi and represent powerful tools to silence gene expression in mammalian cells including genes of viral origin. In this study, we applied siRNAs targeting the VP7 gene of African horse sickness virus (AHSV) that encodes a structural protein required for stable capsid assembly. Using a VP7 expression reporter plasmid and an in vitro model of infection, we show that synthetic siRNA molecules corresponding to the AHSV VP7 gene silenced effectively VP7 protein and mRNA expression, and decreased production of infectious virus particles as evidenced by a reduction in the progeny virion titres when compared to control cells. This work establishes RNAi as a genetic tool for the study of AHSV and offers new possibilities for the analysis of viral genes important for AHSV physiology.

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Acknowledgments

We thank Flip Wege for assistance with cell cultures and virus titrations, and Alan Hall for assistance with fluorescence microscopy. This work was supported by a grant from the National Research Foundation.

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Authors and Affiliations

  1. Department of Microbiology and Plant Pathology, University of Pretoria, Pretoria, 0002, South Africa

    Liesel Stassen & Jacques Theron

  2. Department of Genetics, University of Pretoria, Pretoria, 0002, South Africa

    Henk Huismans

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  1. Liesel Stassen
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  2. Henk Huismans
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  3. Jacques Theron
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Correspondence to Jacques Theron.

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Stassen, L., Huismans, H. & Theron, J. Silencing of African horse sickness virus VP7 protein expression in cultured cells by RNA interference. Virus Genes 35, 777–783 (2007). https://doi.org/10.1007/s11262-007-0162-4

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  • DOI: https://doi.org/10.1007/s11262-007-0162-4

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