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This review delves into the multifaceted antiviral, immunomodulatory, and antitumor physiological roles of type I IFNs, while describing their contribution to the pathogenesis of various disease pathologies including cancer, systemic and organ-specific autoimmunity, neuroinflammation, and atherosclerosis. Genetic determinants influencing activation of type I IFN pathways and therapeutic interventions either targeting or stimulating these pathways in the context of autoimmunity and cancer, respectively, are also discussed. Ultimately, the current understanding of the role of type I IFNs as biomarkers indicative of distinct clinical and serological phenotypes, their correlation with disease activity, their predictive role in therapeutic outcomes across diverse clinical scenarios, as well as the challenges associated with their implementation in clinical practice, are thoroughly addressed. 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