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Synthetic lethality describes a phenomenon where mutation of any one of two non-lethal genes does not affect cell survival, whereas mutation of both genes significantly compromise cell viability. Malignant tumors accumulate a large number of genetic mutations due to errors in DNA replication and repair. Inhibiting another gene that has a synthetic lethal relationship with one of these genes may selectively kill tumor cells without affecting normal cells. In recent years, the principle of synthetic lethality has been successfully applied to the targeted therapy of tumors. This article reviews the basic principles of synthetic lethality, the screening methods for synthetic lethal interactions, the research status of synthetic lethality in the field of DNA damage repair, and the challenges and prospects of synthetic lethality in the development of antitumor drugs.