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<p>Foot-and-mouth disease virus (FMDV) is a highly contagious pathogen that is controlled mainly by the use of inactivated vaccines, but vaccine production is limited by inefficient cell culture systems. The RNA helicase DDX5 has been implicated in viral replication, but its role in FMDV infection remains unclear. Here, we generated a DDX5-knockout PK-15 cell line using CRISPR/Cas9 to investigate its impact on FMDV replication. DDX5 knockout cells exhibited enhanced FMDV replication, with increased viral protein expression, RNA levels, and titers compared to wild-type cells. Meanwhile, RNA sequencing (RNA-seq) analysis indicated that DDX5 knockout suppressed key proinflammatory cytokines (CXCL2/8/14, CCL2/4/5) and impaired IFN-α/β and ISG (ISG15/20, IRF3, IFIT3) responses postinfection. RT-qPCR was performed to determine the expression level of differentially expressed genes, and the results were consistent with the RNA-seq data. Altogether, the results of this study suggest that DDX5 restricts FMDV replication by modulating host innate immunity. The DDX5 knockout cell line provides a useful model for studying FMDV pathogenesis and improving vaccine development.</p>

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CRISPR/Cas9 knockout of DDX5 facilitates foot-and-mouth disease virus replication in PK-15 cells

  • 期刊论文

  • 发表日期：

    2026年3月7日

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  • Jin’en Wu,
  • Yun Zhang,
  • Huichen Guo,
  • Yong Zhang,
  • Yong Zhang

摘要

Foot-and-mouth disease virus (FMDV) is a highly contagious pathogen that is controlled mainly by the use of inactivated vaccines, but vaccine production is limited by inefficient cell culture systems. The RNA helicase DDX5 has been implicated in viral replication, but its role in FMDV infection remains unclear. Here, we generated a DDX5-knockout PK-15 cell line using CRISPR/Cas9 to investigate its impact on FMDV replication. DDX5 knockout cells exhibited enhanced FMDV replication, with increased viral protein expression, RNA levels, and titers compared to wild-type cells. Meanwhile, RNA sequencing (RNA-seq) analysis indicated that DDX5 knockout suppressed key proinflammatory cytokines (CXCL2/8/14, CCL2/4/5) and impaired IFN-α/β and ISG (ISG15/20, IRF3, IFIT3) responses postinfection. RT-qPCR was performed to determine the expression level of differentially expressed genes, and the results were consistent with the RNA-seq data. Altogether, the results of this study suggest that DDX5 restricts FMDV replication by modulating host innate immunity. The DDX5 knockout cell line provides a useful model for studying FMDV pathogenesis and improving vaccine development.