Develop gene-edited chickens with enhanced resistance to highly pathogenic avian influenza (HPAI), a significant threat to poultry health, food security, and public health

The Department of Agriculture Animal and Plant Health Inspection Service awarded The Ohio State University a $1.999 million cooperative agreement on February 2, 2026, to develop gene-edited chickens with enhanced resistance to highly pathogenic avian influenza under the Plant and Animal Disease, Pest Control, and Animal Care program (CFDA 10.025).

The project pursues a host-targeted approach by disrupting four critical genes—ST3GAL1, ST3GAL4, ST3GAL6, and TMPRSS2—that enable viral attachment and activation in chickens. The recipient will create gene-edited chicken lines using an optimized CRISPR-Cas9 platform, validate the gene edits through molecular and biochemical techniques including PCR, DNA sequencing, western blotting, and immunohistochemistry, and assess resistance to HPAI through controlled viral challenge studies conducted in the BSL-3 facility at Ohio State University's Wooster Campus. These challenge studies will evaluate disease progression, viral replication and shedding, survival rates, and tissue responses by comparing gene-edited chickens to non-edited controls.

The cooperative agreement runs through January 31, 2029, with place of performance in Ohio, USA. The project addresses HPAI as a significant threat to poultry health, food security, and public health by preventing infection at the earliest stages of viral entry.

This project takes a novel host-targeted approach by disrupting key genes involved in viral attachment and activation, aiming to prevent infection at the earliest stages of viral entry. To achieve this, the project will focus on three main objectives. First, we will create gene-edited chicken lines by targeting four critical host genes, ST3GAL1, ST3GAL4, ST3GAL6, and TMPRSS2, using our optimized crispr CAS9 platform for poultry species. These genes play a crucial role in the early stages of avian influenza virus (AIV) infection. ST3GAL1, ST3GAL4, and ST3GAL6 encode enzymes that add ?2,3-linked sialic acids to glycoproteins and glycolipids, which act as receptors for the virus in respiratory tissues. TMPRSS2, a host protease, activates the virus by cleaving the hemagglutinin protein, enabling viral entry into cells. Disrupting these genes is expected to significantly reduce AIV susceptibility in chickens. The second objective focuses on validating the success of these gene edits using a range of molecular and biochemical techniques, including PCR, DNA sequencing, western blotting, immunohistochemistry, and glycosylation profiling. This will allow US to confirm the accuracy of the gene knockouts and characterize their functional impact, ensuring the targeted modifications are effective. Finally, we will assess the resistance of these gene-edited chickens to HPAI through controlled viral challenge studies conducted in the BSL-3 facility at the ohio state university wooster campus. These studies will evaluate disease progression, viral replication, shedding, and survival rates, comparing gene-edited chickens to non-edited controls. Histopathology and immunohistochemistry will further examine tissue responses and viral localization. This project will leverage two key areas of expertise: gene editing using crispr CAS9 technology for poultry species and conducting high-security BSL-3 pathogen challenge studies. The expected outcomes include improved poultry health, reduced reliance on culling and vaccination, enhanced biosecurity, and minimized zoonotic risks. Beneficiaries include poultry producers, veterinarians, researchers, and consumers relying on a secure, sustainable poultry supply. No subrecipient activities are planned at this time.