中国兽医科学2025，Vol.55Issue(6)：732-739,8.DOI:10.16656/j.issn.1673-4696.2025.0104
靶向FMDV基因组3B和3Cpro基因CRISPR/Cas13d转基因细胞系的构建及应用
Construction and applications of CRISPR/Cas13d transgenic cell lines targeting foot-and-mouth disease virus 3B and 3Cpro genes
摘要
Abstract
Genome-editing tools offer significant potential for disease resistance.In this study,we investigate the feasibility of CRISPR/Cas13d system in combating the FMDV in vitro.Firstly,we de-signed two guide RNAs(gRNA)targeting the genomic sequences of FMDV non-structural proteins 3B and 3Cpro,and cloned them into the Cas13d expression vector.Cas13d/gRNA transgenic cell lines were produced by transfecting the expression vector into BHK21 cells,followed by puromycin screening.Then,FMDV repli-cation in the transgenic cell lines was detected by RT-qPCR,Western-blot and TCID50 assays.The results demonstrated that the CRISPR/Cas13d system effectively inhibited FMDV replication.The results sug-gest that CRISPR/Cas13d system has great application prospect in disease resistance research on FMDV,providing a theoretical foundation for developing of disease-resistant livestock,and offering a new approach for controlling livestock and poultry diseases.关键词
口蹄疫病毒/CRISPR/Cas13d/家畜抗病/转基因Key words
foot-and-mouth disease virus/CRISPR/Cas13d/livestock disease resistance/geneti-cally modified分类
农业科技引用本文复制引用
郑东东,齐晓兰,杨彪,晏子龙,郑海学,冯涛,徐红伟..靶向FMDV基因组3B和3Cpro基因CRISPR/Cas13d转基因细胞系的构建及应用[J].中国兽医科学,2025,55(6):732-739,8.基金项目
"十四五"广东省揭榜挂帅项目(2024KJ14) （2024KJ14）
科技创新2030重大项目(2023ZD0404301) （2023ZD0404301）
甘肃省联合科研基金重点项目(23JRRA1478) （23JRRA1478）
兰州市科技计划项目(2024-8-12) （2024-8-12）
国家自然科学基金重点项目(32330107) （32330107）
国家重点研发计划项目(2021YFD1800300) （2021YFD1800300）
中央高校基本科研业务费专项资金资助(lzujbky-2022-ct02) （lzujbky-2022-ct02）
国家生猪技术创新中心项目(NCTIP-XD/C03) （NCTIP-XD/C03）
家畜重要疫病诊断研制与产业化项目(22ZD6NA012) （22ZD6NA012）
