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Efficient inhibition of porcine reproductive and respiratory syndrome virus replication by artificial microRNAs targeting the untranslated regions

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Abstract

A robust artificial microRNA (amiRNA) strategy against porcine reproductive and respiratory syndrome virus (PRRSV) was developed by targeting the untranslated regions (UTRs). Six candidate amiRNAs targeting the 5′ or 3′ UTR were used for vector construction, and four effective amiRNAs were selected for further study using a vector transfection/virus infection assay. In cell cultures stably transfected with the four amiRNA vectors, expression of the sequence-specific amiRNAs was confirmed using poly(A)-tailed RT-PCR. After infection with three different PRRSV strains, the viral RNA genome and/or transcript were inhibited by ~90 % (semi-quantitative RT-PCR), and the viral titers were decreased by more than six log CCID50 (viral titration assay) before day 3 postinfection. The potent anti-PRRSV effects lasted for at least 5 days. Sequence analysis showed that the amiRNA antiviral activities were not compromised by the presence of one or two mismatches in their binding targets. This work constitutes a step towards developing a more effective RNAi strategy against PRRSV.

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Acknowledgments

This work was supported by the grant from the Ministry of Agriculture of China (2009ZX08010-019B), the Priority Academic Program Development (PAPD) of Jiangsu Higher Education Institutions, and the Program for Changjiang Scholars and Innovative Research Team in University (IRT0978).

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Authors and Affiliations

  1. College of Veterinary Medicine, Yangzhou University, 48 Wenhui Road, Yangzhou, 225009, Jiangsu, People’s Republic of China

    Bing Xia, Hongqin Song, Yang Chen, Xinyu Zhang, Xiaoli Xia & Huaichang Sun

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  1. Bing Xia
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  2. Hongqin Song
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  3. Yang Chen
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  4. Xinyu Zhang
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  5. Xiaoli Xia
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Correspondence to Huaichang Sun.

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Xia, B., Song, H., Chen, Y. et al. Efficient inhibition of porcine reproductive and respiratory syndrome virus replication by artificial microRNAs targeting the untranslated regions. Arch Virol 158, 55–61 (2013). https://doi.org/10.1007/s00705-012-1455-z

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  • DOI: https://doi.org/10.1007/s00705-012-1455-z

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