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CRISPR/Cas9-mediated multiple single guide RNAs potently abrogate pseudorabies virus replication

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Abstract

Pseudorabies virus (PRV) is a swine herpesvirus that causes significant morbidity and mortality in swine populations and has caused huge economic losses in the worldwide swine industry. Currently, there is no effective antiviral drug in clinical use for PRV infection; it is also difficult to eliminate PRV from infected swine. In our study, we set out to combat these swine herpesvirus infections by exploiting the CRISPR/Cas9 system. We designed 75 single guide RNAs (sgRNA) by targeting both essential and non-essential genes across the genome of PRV. We applied a firefly luciferase-tagged reporter PRV virus for high-throughput sgRNA screening and found that most of the sgRNAs significantly inhibited PRV replication. More importantly, using a transfection assay, we demonstrated that simultaneous targeting of PRV with multiple sgRNAs completely abolished the production of infectious viruses in cells. These data suggest that CRISPR/Cas9 could be a novel therapeutic agent against PRV in the future.

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Acknowledgements

This study was supported by grants from the National Program on Key Research Project (2016YFD0500100).

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Author notes

  1. Yan-Dong Tang, Ji-Ting Liu and Tong-Yun Wang contributed equally to this study.

Authors and Affiliations

  1. State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, No. 678 HaPing Road, XiangFang, Harbin, 150069, China

    Yan-Dong Tang, Ji-Ting Liu, Tong-Yun Wang, Ming-Xia Sun, Zhi-Jun Tian & Xue-Hui Cai

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  1. Yan-Dong Tang
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  2. Ji-Ting Liu
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  3. Tong-Yun Wang
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  4. Ming-Xia Sun
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Correspondence to Xue-Hui Cai.

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Handling Editor: Günther Keil.

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Tang, YD., Liu, JT., Wang, TY. et al. CRISPR/Cas9-mediated multiple single guide RNAs potently abrogate pseudorabies virus replication. Arch Virol 162, 3881–3886 (2017). https://doi.org/10.1007/s00705-017-3553-4

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  • DOI: https://doi.org/10.1007/s00705-017-3553-4

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