Methodological Landscape of DNA Damage Response Detection: From Conventional Assays to Future Innovations

All living organisms possess a DNA damage response (DDR) that is important for genetic evolution. Cells have developed comprehensive mechanisms for addressing DNA damage, collectively called the DNA damage response and repair. External environmental stress continuously disrupts genomic integrity and triggers various pathological changes. The failure of the DDR network often drives cell carcinogenesis, and its core components not only serve as biological markers for disease monitoring but also represent highly promising molecular targets for targeted therapy. Therefore, there is a high level of interest in exploring DDR-related biomarkers as cutting-edge therapeutic regimens and developing highly sensitive tools for DDR diagnosis. These methods should assess the rate of damage occurrence and distinguish when repair pathways are activated. These kinds of advances are key to preserving genetic stability and detecting and preventing diseases early. Here, we provide a broad summary of recent advances in DDR detection technologies, with a particular focus on the complementarity between different techniques. We have also summarized current technological bottlenecks, future research paradigms, and clinical translation pathways. The insights presented in this review will contribute to the development of multidisciplinary integrated DDR detection technologies, promote the establishment of DDR biomarker detection systems, and provide crucial methodological references for targeted drug development, efficacy evaluation, and resistance mechanism research targeting the DDR pathway.