Background: Interferon Regulatory Factor 8 (IRF8) is a transcription factor integral to the immune system, orchestrating various aspects of immune response regulation. Previous studies have established IRF8’s role in myeloid cell differentiation and have linked it to several immune disorders. However, the full scope of its genetic variability, functional domains, and interaction networks remained insufficiently characterized. Objective: This study aimed to conduct a comprehensive analysis of the IRF8 gene and protein to explore its structure, function, regulatory mechanisms, and potential as a therapeutic target. Methods: We utilized a combination of bioinformatics tools to map the genomic location of IRF8, predict its protein structure, and identify functional domains. Gene and protein expressions were profiled using public databases, and post-translational modifications were predicted through specialized software. Protein-protein interaction networks were examined using the STRING database, and potential drug-binding pockets were identified via the DoGSite scorer. Splice variants and polymorphisms were assessed for clinical significance, and subcellular localization was predicted using several computational tools. Results: The IRF8 gene was confirmed to encode a protein of 426 amino acids, with no methylation sites predicted. Its expression was notably higher in lymphoid tissues, and it exhibited extensive nuclear localization. We identified 11 splice variants and 11,168 single nucleotide polymorphisms (SNPs), with varying implications for pathogenicity. The protein interactome analysis revealed a central role in immune signaling pathways. Moreover, several high-affinity drug-binding pockets were identified, suggesting IRF8 as a promising therapeutic target. Conclusion: The study provided detailed insights into the IRF8 gene and protein, suggesting significant regulatory complexity and highlighting its potential in disease pathogenesis and therapy. Our findings support the role of IRF8 as a pivotal element in immune response and offer a foundation for the development of novel therapeutic approaches.