pAPN和NEU3的敲除对猪传染性胃肠炎病毒(TGEV)侵染的影响  

Effects of pAPN and NEU3 knockout on TGEV infection

作　　者：李兆龙[1,2] 丰志华 张冰晨 方舟 梁旺旺 陈文志 LI Zhaolong;FENG Zhihua;ZHANG Bingchen;FANG Zhou;LIANG Wangwang;CHEN Wenzhi(Institute of Animal Husbandry and Veterinary Medicine,Fujian Academy of Agricultural Sciences,Fuzhou 350013,China;Biomedical Research Center of South China,Fujian Normal University,Fuzhou350117,China)

机构地区：[1]福建省农业科学院畜牧兽医研究所,福建福州350013 [2]福建师范大学南方生物医学研究中心,福建福州350013

出　　处：《中国兽医学报》2022年第10期1942-1948,1981,共8页Chinese Journal of Veterinary Science

摘　　要：氨基肽酶(pAPN)是猪传染性胃肠炎病毒(TGEV)侵染的主要受体,为了验证pAPN和唾液酸神经氨酸酶(NEU3)双基因对TGEV入侵机制的影响,应用CRISPR-Cas9基因编辑技术,敲除ST细胞的2个基因pAPN和NEU3。经过病毒感染试验,测定敲除2个目标基因pAPN和NEU3的ST细胞,对病毒的侵染变化以及病毒拷贝数的变化和细胞病变改善,同时监测病毒侵染后纤连蛋白的变化。结果表明,与对照组相比,敲除2个目标基因pAPN和NEU3的ST细胞,明显减轻TGEV侵染引起的细胞病变,TGEV的拷贝数也明显出现下降。此外,同样滴度的TGEV侵染pAPN和NEU3双基因敲除ST细胞后,诱导ST细胞的免疫应答物IFN-β的mRNA水平明显低于野生型ST细胞组。pAPN和NEU3双基因的敲除,明显降低了ST细胞中TGEV的拷贝数,同时也减轻病毒引起的细胞病变,结果证实细胞中的pAPN和NEU3双基因可作为将来养猪生产中抗病毒治疗及抗病品种选育的靶基因。The aminopeptidase(pAPN) is the main receptor for TGEV infection.This study aims to verify the effect of pAPN and sialic acid neuraminidase(NEU3) on TGEV virus infection.pAPN and sialic acid neuraminidase(NEU3) were knockout using CRISPR gene editing techniques.The virus infection test was used to determine the infection changes of ST cells with pAPN and NEU3 target genes knocked out, the changes of virus copy number and the improvement of cytopathic effect.At the same time, the changes of fibronectin after virus infection were monitored.The results showed that compared with the control group, ST cells with pAPN and NEU3 target genes knocked out significantly attenuated TGEV infection-induced cytopathies.TGEV copy number also decreased significantly.In addition, pAPN and NEU3 double knockout ST cells were infected with the same titer of TGEV,the results showed that the mRNA level of immune responders was significantly lower than that of wild-type ST cells.In conclusion, pAPN and sialic acid NEU3 can inhibit TGEV virus infection, reduce viral-induced cytopathies.This result confirms that pAPN and NEU3 genes in cells can be used as target genes for antiviral therapy and breeding of resistant varieties in pig production in the future.

关 键 词：氨基肽酶 唾液酸神经氨酸酶 猪传染性胃肠炎病毒 病毒的拷贝数 

分 类 号：S852.65[农业科学—基础兽医学]