中国兽医科学2026，Vol.56Issue(1)：27-32,6.DOI:10.16656/j.issn.1673-4696.2025.0231
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
Establishment of a proline oligopeptidase gene knockout PK-15 cell line and its impact on foot-and-mouth disease virus replication
摘要
Abstract
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.关键词
脯氨酰寡肽酶/CRISPR/Cas9/基因敲除/口蹄疫病毒/病毒增殖Key words
prolyl oligopeptidase/CRISPR/Cas9/gene knockout/foot-and-mouth disease virus/vi-ral replication分类
农业科技引用本文复制引用
卢炳州,郑海学,郭建宏,茹毅,郝荣增,李建斌,王姿逸,杨洋,赵陇和,李亚军,刘华南,李明桂,马坤..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[J].中国兽医科学,2026,56(1):27-32,6.基金项目
国家自然科学基金项目(32372990) （32372990）
国家重点研发计划项目(2024YFD1800501) （2024YFD1800501）
甘肃省自然科学基金重点项目(23YFNA0011) （23YFNA0011）
甘肃省自然科学基金项目(23JRRA549) （23JRRA549）
