华南农业大学学报2026，Vol.47Issue(3)：431-441,11.DOI:10.7671/j.issn.1001-411X.202508001
新型CD163截短型基因编辑猪的制备及抗猪繁殖与呼吸综合征效果评估
Establishment of CD163-truncated gene-edited pigs and evaluation of their anti-PRRSV ability
摘要
Abstract
[Objective]This study aims to generate Large White pigs with a five-amino acid deletion(Δ525-529,AHFGE)in the SRCR5 domain of CD163 using CRISPR/Cas9 gene editing combined with somatic cell nuclear transfer,and evaluate their resistance to porcine reproductive and respiratory syndrome virus(PRRSV)infection.[Method]In the cellular experiment,HEK-293T cells were transfected with plasmids expressing wild-type CD163(pCMV-pCD163-neo)or truncated CD163(pCMV-pCD163-ΔAHFGE).Twenty-four hours later,the cells were inoculated with the PRRSV JXA1 strain.Viral infection was assessed 24 hours post-inoculation by fluorescence observation and immunofluorescence staining.In the animal experiment,a total of 20 pigs were included and divided into three groups:the AHFGE precise deletion group(n=6),the wild-type challenged control group(n=9),and the wild-type unchallenged blank control group(n=5).Pigs in the AHFGE precise deletion group and the wild-type challenged control group received an intramuscular injection of 2 mL PRRSV JXA1 strain suspension(virus titer was 5×104 TCID50/mL).All pigs were monitored for 21 consecutive days post-challenge.The main observed indicators included clinical symptoms,rectal temperature changes,serum PRRSV load,and specific antibody levels.[Result]Gene-edited Large White pigs with the five-amino acid deletion Δ525-529(AHFGE)in the SRCR5 domain of CD163 were successfully generated.The gene-edited pigs showed normal overall growth,with no significant phenotypic abnormalities observed.Animal challenge experiments showed that the gene-edited pigs had enhanced resistance to the highly pathogenic PRRSV JXA1 strain.They displayed milder disease phenotypes and had obviously lower in vivo viral loads,antibody levels,as well as mortality compared with wild-type controls of the same breed.[Conclusion]The AHFGE motif is the critical amino acid residues mediating PRRSV infection.The gene-edited pigs generated in this study can serve as breeding materials for cultivating PRRSV-resistant pig populations,laying a foundation for evaluating their breeding value and application potential.关键词
猪/CD163截短型/基因编辑/AHFGE/PRRSV/抗病育种Key words
Pig/CD163-truncated/Gene editing/AHFGE/PRRSV/Antiviral breeding分类
农业科技引用本文复制引用
谭嘉泳,张健,杨晓辉,钟海文,杨化强,蔡更元..新型CD163截短型基因编辑猪的制备及抗猪繁殖与呼吸综合征效果评估[J].华南农业大学学报,2026,47(3):431-441,11.基金项目
广东省种业振兴行动项目(2024-XPY-00-015) （2024-XPY-00-015）
科技创新2030重大项目(2023ZD0404303) （2023ZD0404303）
国家自然科学基金(32372874) （32372874）
