猪肾上皮细胞Ⅰ型干扰素受体1敲除对伪狂犬病毒复制的影响

Effect of porcine kidney epithelial cells type Ⅰ interferon receptor 1 knockdown on pseudorabies virus replication

目的探讨Ⅰ型干扰素受体1 (IFNAR1)对伪狂犬病病毒(PRV)复制的影响。方法以慢病毒介导的CRISPR/Cas9基因编辑技术,构建猪肾上皮细胞(PK15)敲除IFNAR1基因的稳定细胞系。运用细胞活力检测、荧光观察、流式细胞术检测、滴度测定、Real-time PCR等技术进行IFNAR1功能验证。结果随着PRV感染时间延长,敲除IFNAR1基因可以显著促进PRV-TK mRNA的转录,PRV-gE蛋白的翻译以及子代病毒的毒力。结论IFNAR1在抑制PRV增殖中发挥重要作用。

Objective To investigate the effect of type Ⅰ interferon receptor 1( IFNAR1) on pseudorabies virus( PRV) replication. Methods A stable cell line in which porcine kidney epithelial cells( PK15) knocked out the IFNAR1 gene was constructed using a lentiviral-mediated CRISPR/Cas9( clustered regularly interspaced short palindromic repeats,CRISPR) gene editing technique. The function of IFNAR1 was verified by cell viability assay,fluorescence observation,flow cytometry detection,titer determination,and Real-time PCR. Results With the prolongation of PRV infection,knocking out the IFNAR1 gene can significantly promote transcription of PRV-TK mRNA,translation of PRV-gE protein,and virulence of progeny virus. Conclusion IFNAR1 plays an important role in inhibiting the proliferation of PRV.