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ARS Home » Plains Area » Clay Center, Nebraska » U.S. Meat Animal Research Center » Animal Health Genomics » Research » Publications at this Location » Publication #426072
Research Project: Strategies to Control Respiratory Diseases of Cattle

Location: Animal Health Genomics

Title: CD46 gene-editing confers ex vivo BVDV resistance in fibroblasts from cloned Angus calves

Author
item Workman, Aspen
item Heaton, Michael
item VANDER LEY, BRIAN - University Of Nebraska

Submitted to: Viruses
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/24/2025
Publication Date: 5/29/2025
Citation: Workman, A.M., Heaton, M.P., Vander Ley, B.L. 2025. CD46 gene-editing confers ex vivo BVDV resistance in fibroblasts from cloned Angus calves. Viruses. 17(6). Article 775. https://doi.org/10.3390/v17060775.
DOI: https://doi.org/10.3390/v17060775

Interpretive Summary: Bovine Viral Diarrhea Virus (BVDV) remains a leading global cause of disease and reproductive losses in cattle. Our previous work used CRISPR/Cas9 gene-editing technology to produce BVDV resistance in a cloned Gir breed heifer calf. Here, we show that the same genetic modification significantly protects cells from gene-edited cloned American Angus calves against BVDV infection in the laboratory (ex vivo). The consistent cellular resistance to the virus underscores the strong potential for broad application of this approach across popular cattle breeds. If successful, implementation of this gene-editing strategy could significantly reduce the economic burden and animal welfare concerns associated with BVDV, leading to healthier and more productive cattle herds.

Technical Abstract: A previous study demonstrated that a 19-nucleotide edit, encoding a six amino acid substitution in the bovine CD46 gene, dramatically reduced bovine viral diarrhea virus (BVDV) susceptibility in a cloned Gir (Bos indicus) heifer. The present study aimed to replicate this result in American Angus (Bos taurus) using genetically matched controls and larger sample sizes. CRISPR/Cas9-mediated homology-directed repair introduced the identical CD46 edit, encoding the A82LPTFS amino acid sequence, into exon 2 of CD46 in primary Angus fibroblasts. Thirty-three cloned embryos (22 CD46-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 CD46 editing in BVDV control. Continued advancements in cloning technology will enhance the potential of gene-editing for producing disease-resistant livestock.