Construction of Interferon Regulatory Factor Knockdown Cell Line and Its Effect on Pseudorabies Virus Proliferation
To elucidate the impact of interferon regulatory factors(IRFs)on the proliferation of porcine pseudorabies virus(PRV),this study utilized shRNA technology to establish a PK15 cell line with the targeted knockdown of IRF1-9 genes.The knockdown efficiency was verified,and the proliferation of PRV subsequent to the knockdown was assessed using flow cytometry and titration assays.Additionally,the mRNA expression levels of PRV-gB,IFN-β,ISG-15,and IL-6 in PRV-infected cells were quantified by reverse transcription quantitative polymerase chain reaction(RT-qPCR).The expression of the gB protein in PRV-infected cells was analyzed by Western blotting.The knockdown efficiency assay demonstrated a significant reduction in the expression of IRF1-9 mRNA in PK15 cells.The results from flow cytometry and titration assays indicated that the knockdown of IRF1-9 genes facilitated the proliferation of PRV.Furthermore,RT-qPCR and Western blot results revealed a significant increase in the mRNA and protein expression levels of PRV-gB following the knockdown of IRF1-9 genes.The analysis of mRNA expression levels of cellular inflammatory factors showed that the knockdown of IRF1-9 inhibited the PRV-induced expression of IFN-β,ISG-15,and IL-6 at the mRNA level.In conclusion,these findings suggest that IRF1-9 serves as a host restriction factor,limiting the replication of PRV within PK-15 cells.
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维普
