Cyclic GMP-AMP synthase (cGAS) is a cytosolic DNA sensor that initiates innate immune responses. DNA-bound cGAS produces cyclic GMP-AMP (cGAMP), which activates stimulator of interferon genes (STING) to induce inflammatory cytokines and other immune mediators. cGAS detects DNA without sequence specificity and responds to both cytosolic foreign DNA from pathogens and self-DNA leaked into the cytosol due to genome instability or cellular damage. Because of the diverse sources of cytosolic DNA, the cGAS-STING pathway plays a critical role during infection, autoimmune diseases, and senescence. Moreover, cGAS detects tumor-derived DNA and stimulates endogenous antitumor immunity. Thus, the cGAS-STING pathway is a promising target for cancer immunotherapy. Here, we review the role of the cGAS-STING pathway in various diseases and highlight various approaches targeting the cGAS-STING pathway for cancer therapy.