中国兽医科学2025，Vol.55Issue(7)：862-869,8.DOI:10.16656/j.issn.1673-4696.2025.0122
CISD2敲除细胞系的构建及其对口蹄疫病毒复制的影响
Construction of cell line with CISD2 gene knockout and its impact on the replication of foot-and-mouth disease virus
摘要
Abstract
The aim of this study was to preliminarily investigate the effect of cellular iron-sulfur protein 2(CISD2)on the replication of foot-and-mouth disease virus(FMDV).A CISD2 knockdown porcine kidney cell line(PK-15)was successfully constructed using CRISPR/Cas9 gene editing method,and the knockdown of CISD2 gene was identified by sequencing and Western-blot,as well as indirect immunofluo-rescence assay,Western-blot,and RT-qPCR to compare the replication of FMDV in the CISD2 knockdown cell line and the wild type,and statistically analyze the effect of CISD2 on FMDV replication.The results showed that sequencing and Western-blot confirmed the successful construction of CISD2 knockout mono-clonal cell lines,and indirect immunofluorescence assay,Western-blot,and RT-qPCR showed that the replication level of FMDV in CISD2 knockout cells was significantly higher than that in wild-type cells,and the study successfully utilized the CRISPR/Cas9 system to The study successfully constructed a CISD2 knockout cell line using the CRISPR/Cas9 system and found that FMDV replication was significantly promoted in this cell.The above results provide materials and data for further investigation of the function of CISD2 and its role in regulating FMDV and other viral infections.关键词
口蹄疫病毒/CRISPR/Cas9/CISD2基因/细胞系Key words
foot-and-mouth disease virus/CRISPR/Cas9/CISD2/cell line分类
农业科技引用本文复制引用
陈治彤,张伟,蔡建涛,吕岳芹,刘泓屹,曹伟军,顾有方,郑海学,杨帆..CISD2敲除细胞系的构建及其对口蹄疫病毒复制的影响[J].中国兽医科学,2025,55(7):862-869,8.基金项目
国家自然科学基金项目(32072831,32102639) （32072831,32102639）
国家重点研发计划项目(2021YFD1800300) （2021YFD1800300）
甘肃省杰出青年基金项目(21JR7RA026) （21JR7RA026）
兰州大学中央高校基本科研业务费专项(lzujbky-2022-ey20) （lzujbky-2022-ey20）
甘肃省科技重大专项计划项目(22ZD6NA001) （22ZD6NA001）
