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Resistance to pseudorabies virus by knockout of nectin1/2 in pig cells

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Abstract

Pseudorabies virus (PRV) is a pig pathogen that causes substantial economic losses to the pig industry. Infection of host cells by PRV is mediated by the membrane proteins nectin1 and nectin2, which are presumed to be receptors for PRV infection. Here, we generated nectin1/2 knockout (KO) cells with the aim of establishing a PRV-resistant cell model. Nectin1 and 2 were ablated in PK15 cells by CRISPR/Cas9-mediated gene targeting. PRV infection in either nectin1 or nectin2 KO cells showed a significant reduction in viral growth compared with wild-type (WT) cells. We further simultaneously deleted nectin1 and nectin2 in PK15 cells and found that double KO cells showed no further increase in resistance to PRV compared with single gene-KO cells, despite being more resistant than WT. By investigating the cell entry steps of PRV infection, we found that nectin1 or/and nectin2 KO did not greatly affect virus attachment or internalization to cells but blocked cell-to-cell spread. Our results demonstrate that KO of either nectin1 or nectin2 confers PRV resistance to PK15 cells. This strategy could be applied to establish PRV-resistant pigs with nectin1/2 modifications to benefit the pig industry.

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Acknowledgements

This work was supported in part by a grant from the National Natural Science Foundation of China (31772555).

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Authors and Affiliations

  1. National Engineering Research Center for Breeding Swine Industry, College of Animal Science, South China Agricultural University, Guangzhou, China

    Yaoqiang Huang, Zicong Li, Changxu Song, Zhenfang Wu & Huaqiang Yang

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  1. Yaoqiang Huang
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  2. Zicong Li
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Correspondence to Zhenfang Wu or Huaqiang Yang.

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Huang, Y., Li, Z., Song, C. et al. Resistance to pseudorabies virus by knockout of nectin1/2 in pig cells. Arch Virol 165, 2837–2846 (2020). https://doi.org/10.1007/s00705-020-04833-x

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  • DOI: https://doi.org/10.1007/s00705-020-04833-x

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