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Published online ahead of print on 4 March 2009 as doi:10.1099/vir.0.010520-0
Journal of General Virology 2009;90:1417.

A more recent version of this article appeared on June 1, 2009 J Gen Virol (2009), DOI 10.1099/vir.0.010520-0
© 2009 Society for General Microbiology
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Effective inhibition of infectious bursal disease virus replication by recombinant avian adeno-associated virus-delivered microRNAs

Huaichang Sun1, Yongjuan Wang, Pengpeng Shen, Xinyu Zhang and Xiaoli Xia

Yangzhou University

1 E-mail: sunh{at}yzu.edu.cn

RNA interference (RNAi) is a novel anti-viral strategy against a variety of viral infections. Infectious bursal disease virus (IBDV) causes an economically important disease in young chickens. In this study, we demonstrated efficient inhibition of IBDV replication by recombinant avian adeno-associated virus (rAAAV)-delivered anti-VP1 and anti-VP2 microRNAs (miRNAs). In the viral vector-transduced cells, the sequence-specific miRNA expression was detected by poly(A)-tailed reverse transcriptase PCR (RT-PCR). Reporter assays using the pVP2-EGFP vector showed significant and long-lasting inhibition of the VP2-EGFP expression in cells transduced with the anti-VP2 miRNA-expressing rAAAV-RFPmiVP2E, but not with the control miRNA-expressing rAAAV-RFPmiVP2con or anti-VP1 miRNA-expressing rAAAV-RFPmiVP1. Semi-quantitative RT-PCR and/or virus titration assays showed significant inhibitory effect on homologous IBDV replication in cells transduced with rAAAV-RFPmiVP1 or rAAAV-RFPmiVP2E. For two heterologous IBDV isolates, transduction with rAAAV-RFPmiVP1 led to slightly weaker but similar inhibitory effects, whereas transduction with rAAAV-RFPmiVP2E resulted in significantly weaker and different inhibitory effects. These results suggest that rAAAV could act as an efficient vector for miRNA delivery into avian cells and VP1 was the more suitable target for interfering IBDV replication using RNAi technology.

Received 23 January 2009; accepted 29 January 2009.




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