First gene-edited calf with reduced susceptibility to a major viral pathogen

Authors

• Aspen M. Workman, USDA, ARS, U.S. Meat Animal Research CenterFollow
• Michael P. Heaton, USDA, ARS, U.S. Meat Animal Research CenterFollow
• Brian L. Vander Ley, University of Nebraska-LincolnFollow
• Dennis A. Webster, Recombinetics Inc., Eagan, MN
• Luke Sherry, Recombinetics Inc., Eagan, MN
• Jonathan R. Bostrom, Recombinetics Inc., Eagan, MN
• Sabrina Larson, Acceligen Inc., Eagan, MN
• Theodore S. Kalbfleisch, University of Kentucky
• Gregory P. Harhay, US Meat Animal Research Center, Clay Center, NebraskaFollow
• Erin E. Jobman, University of Nebraska-Lincoln
• Daniel F. Carlson, Recombinetics Inc., Eagan, MN
• Tad S. Sonstegard, Acceligen Inc., Eagan, MN

Document Type

Article

Date of this Version

2023

Citation

PNAS Nexus, 2023, 2, 1–14

https://doi.org/10.1093/pnasnexus/pgad125

Comments

U.S. government work

Abstract

Bovine viral diarrhea virus (BVDV) is one of the most important viruses affecting the health and well-being of bovine species throughout the world. Here, we used CRISPR-mediated homology-directed repair and somatic cell nuclear transfer to produce a live calf with a six amino acid substitution in the BVDV binding domain of bovine CD46. The result was a gene-edited calf with dramatically reduced susceptibility to infection as measured by reduced clinical signs and the lack of viral infection in white blood cells. The edited calf has no off-target edits and appears normal and healthy at 20 months of age without obvious adverse effects from the on-target edit. This precision bred, proof-of-concept animal provides the first evidence that intentional genome alterations in the CD46 gene may reduce the burden of BVDV-associated diseases in cattle and is consistent with our stepwise, in vitro and ex vivo experiments with cell lines and matched fetal clones.