Workman Aspen M, Heaton Michael P, Vander Ley Brian L
US Meat Animal Research Center, USDA, Agricultural Research Service, Clay Center, NE 68933, USA.
Great Plains Veterinary Educational Center, University of Nebraska-Lincoln, Clay Center, NE 68933, USA.
Viruses. 2025 May 29;17(6):775. doi: 10.3390/v17060775.
A previous study demonstrated that a 19-nucleotide edit, encoding a six amino acid substitution in the bovine gene, dramatically reduced bovine viral diarrhea virus (BVDV) susceptibility in a cloned Gir () heifer. The present study aimed to replicate this result in American Angus () using genetically matched controls and larger sample sizes. CRISPR/Cas9-mediated homology-directed repair introduced the identical edit, encoding the ALPTFS amino acid sequence, into exon 2 of in primary Angus fibroblasts. Thirty-three cloned embryos (22 -edited and 11 unedited) were transferred to recipient cows. However, all pregnancies resulted in pre- and perinatal losses due to cloning-related abnormalities, preventing in vivo BVDV challenge. Consequently, ex vivo BVDV susceptibility assays were performed on primary fibroblast cell lines rescued from deceased cloned Angus calves. Infection studies revealed significantly reduced susceptibility in the edited lines, comparable to the resistance previously observed from the edited Gir heifer. These studies extend the applicability of this finding from Gir to the most common US beef breed, Angus, suggesting the potential for broad application of editing in BVDV control. Continued advancements in cloning technology will enhance the potential of gene-editing for producing disease-resistant livestock.
先前的一项研究表明，在牛基因中编码六个氨基酸替换的19个核苷酸编辑，显著降低了克隆的吉尔（Gir）小母牛对牛病毒性腹泻病毒（BVDV）的易感性。本研究旨在使用基因匹配的对照和更大的样本量在美国安格斯（Angus）牛中复制这一结果。CRISPR/Cas9介导的同源定向修复将编码ALPTFS氨基酸序列的相同编辑引入原代安格斯成纤维细胞的外显子2中。33个克隆胚胎（22个经过编辑，11个未经过编辑）被移植到受体母牛体内。然而，由于克隆相关异常，所有妊娠均导致产前和围产期损失，从而无法进行体内BVDV攻击。因此，对从死亡的克隆安格斯犊牛中挽救的原代成纤维细胞系进行了体外BVDV易感性测定。感染研究表明，编辑后的细胞系易感性显著降低，与先前在编辑后的吉尔小母牛中观察到的抗性相当。这些研究将这一发现的适用性从吉尔牛扩展到了美国最常见的肉牛品种安格斯牛，表明基因编辑在BVDV控制中具有广泛应用的潜力。克隆技术的持续进步将提高基因编辑在生产抗病家畜方面的潜力。
