State Key Laboratory of Animal Nutrition and Key Laboratory of Animal Genetics, Breeding and Reproduction of Ministry of Agriculture and Rural Affairs of China, Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.
State Key Laboratory of Agricultural Microbiology and Key Laboratory of Preventive Veterinary Medicine in Hubei Province, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan, China.
Elife. 2020 Sep 2;9:e57132. doi: 10.7554/eLife.57132.
Porcine reproductive and respiratory syndrome virus (PRRSV) and transmissible gastroenteritis virus (TGEV) are two highly infectious and lethal viruses causing major economic losses to pig production. Here, we report generation of double-gene-knockout (DKO) pigs harboring edited knockout alleles for known receptor proteins CD163 and pAPN and show that DKO pigs are completely resistant to genotype 2 PRRSV and TGEV. We found no differences in meat-production or reproductive-performance traits between wild-type and DKO pigs, but detected increased iron in DKO muscle. Additional infection challenge experiments showed that DKO pigs exhibited decreased susceptibility to porcine deltacoronavirus (PDCoV), thus offering unprecedented in vivo evidence of pAPN as one of PDCoV receptors. Beyond showing that multiple gene edits can be combined in a livestock animal to achieve simultaneous resistance to two major viruses, our study introduces a valuable model for investigating infection mechanisms of porcine pathogenic viruses that exploit pAPN or CD163 for entry.
猪繁殖与呼吸综合征病毒（PRRSV）和传染性胃肠炎病毒（TGEV）是两种高度传染性和致命性的病毒，给养猪业造成了重大经济损失。在这里，我们报告了携带已知受体蛋白 CD163 和 pAPN 编辑敲除等位基因的双基因敲除（DKO）猪的产生，并表明 DKO 猪对 2 型 PRRSV 和 TGEV 完全具有抗性。我们发现野生型和 DKO 猪在肉质生产或繁殖性能特征上没有差异，但在 DKO 肌肉中检测到铁含量增加。额外的感染挑战实验表明，DKO 猪对猪德尔塔冠状病毒（PDCoV）的敏感性降低，从而为 pAPN 作为 PDCoV 受体之一提供了前所未有的体内证据。除了表明可以在牲畜动物中组合多种基因编辑以实现对两种主要病毒的同时抗性之外，我们的研究还为研究利用 pAPN 或 CD163 进入的猪致病性病毒的感染机制引入了一个有价值的模型。
