Division of Animal Science, University of Missouri, Randall Prather, 920 East Campus Drive, Columbia, MO, 65211, USA.
Department of Diagnostic Medicine and Pathobiology, Kansas State University, Manhattan, KS, 66506, USA.
Transgenic Res. 2019 Feb;28(1):21-32. doi: 10.1007/s11248-018-0100-3. Epub 2018 Oct 12.
The alphacoronaviruses, transmissible gastroenteritis virus (TGEV) and Porcine epidemic diarrhea virus (PEDV) are sources of high morbidity and mortality in neonatal pigs, a consequence of dehydration caused by the infection and necrosis of enterocytes. The biological relevance of amino peptidase N (ANPEP) as a putative receptor for TGEV and PEDV in pigs was evaluated by using CRISPR/Cas9 to edit exon 2 of ANPEP resulting in a premature stop codon. Knockout pigs possessing the null ANPEP phenotype and age matched wild type pigs were challenged with either PEDV or TGEV. Fecal swabs were collected daily from each animal beginning 1 day prior to challenge with PEDV until the termination of the study. The presence of virus nucleic acid was determined by PCR. ANPEP null pigs did not support infection with TGEV, but retained susceptibility to infection with PEDV. Immunohistochemistry confirmed the presence of PEDV reactivity and absence of TGEV reactivity in the enterocytes lining the ileum in ANPEP null pigs. The different receptor requirements for TGEV and PEDV have important implications in the development of new genetic tools for the control of enteric disease in pigs.
α冠状病毒、传染性胃肠炎病毒（TGEV）和猪流行性腹泻病毒（PEDV）是导致新生仔猪高发病率和高死亡率的病原体，其会引起仔猪感染和肠细胞坏死导致的脱水。通过使用 CRISPR/Cas9 编辑 ANPEP 基因的外显子 2 产生提前终止密码子，评估天冬氨酰肽酶 N（ANPEP）作为 TGEV 和 PEDV 在猪中潜在受体的生物学相关性。敲除 ANPEP 基因的纯合子缺失表型的猪与年龄匹配的野生型猪一起接受 PEDV 或 TGEV 挑战。从 PEDV 攻毒前 1 天开始，每天从每只动物收集粪便拭子，直到研究结束。通过 PCR 确定病毒核酸的存在。ANPEP 基因缺失的猪不支持 TGEV 感染，但仍然易感染 PEDV。免疫组化证实了 ANPEP 基因缺失的猪回肠上皮细胞中存在 PEDV 反应，而不存在 TGEV 反应。TGEV 和 PEDV 对受体的不同要求对开发用于控制猪肠道疾病的新型遗传工具具有重要意义。
