The chicken is an example of efficient intensive animal agricultural production, serving as a primary source of animal protein globally. However, the industry faces significant challenges from infectious diseases, notably avian influenza (AI), which has serious welfare and economic implications. The increasing number of AI outbreaks in poultry farms globally, along with sporadic spillover events in mammals, including humans, underscores the need for innovative strategies to mitigate future production losses and prevent potential human pandemics. A potential solution may be through introgression of the AIV-resilience trait into chickens through gene editing techniques. . However, the successful implementation of gene editing relies on our ability to identify the host-factors critical for AIV in chickens. Prior studies employing Genome-wide CRISPR KO (GeCKO) screening have identified numerous host factors for influenza viruses in human cells. To our knowledge, no similar studies have been reported in chicken cells. To address this gap, we are currently performing a GeCKO screen in chicken fibroblast DF1 cells to identify the host- factors required for AIV infection. This involves the development of a GeCKO cell library, wherein cells are targeted by gRNAs at the genome scale. Next, we will perform an H7N7 AI infection survival screen on GeCKO library cells, during which multiple rounds of AI infection will be performed, with virus-induced cell death serving as the readout. Following NGS and bioinformatics analyses, the highly enriched genes from the survival screen will be validated through a range of functional analyses to dissect the mechanisms by which these genes confer resistance to infection. The outcomes of GeCKO screening may provide new insights into the AI-host interface across the viral life cycle, deepen our understanding of host-directed approaches, and aid in the discovery of novel gene-editing targets to address AIV in chickens.