POP基因敲除PK-15细胞系的建立及其对口蹄疫病毒增殖的影响OA

To investigate the role of prolyl oligopeptidase(POP)in regulating foot-and-mouth disease virus(FMDV)replication,CRISPR/Cas9 gene editing was employed to generate a POP-deficient porcine kidney epithelial cell line(PK-15).Lentiviral transduction,puromycin selection,and limiting dilution cloning were utilized to establish the POP-knockout PK-15 cell line(POP-KO),with wild-type(WT)PK-15 cells serving as controls.Post-FMDV infection,viral replication kinetics were quantita-tively analyzed through real-time quantitative reverse transcription PCR(RT-qPCR)for viral RNA quan-tification,Western-blot analysis of structural protein VP1 expression,and 50％tissue culture in-fectious dose(TCID50)assays for viral titer determination.Comparative analyses demonstrated that POP-KO cells exhibited reduction in VP1 protein levels compared to WT-PK-15 cells(P＜0.001),accompa-nied by a 1.24 lg TCID50/mL decrease in viral titers(P＜0.001).Quantitative PCR revealed suppression of FMDV RNA copy numbers in POP-KO cells(P＜0.001).This study successfully established a POP-defi-cient PK-15 model,demonstrating that POP deficiency potently inhibits FMDV replication kinetics and confirming POP as an essential host factor facilitating FMDV propagation.