Abstract 1480: Unlocking DNA repair mechanisms with dynamic single-molecule analysis

Abstract The maintenance of genomic information through processes such as DNA replication, recombination, and repair is critical to cellular function. Errors in these underlying mechanisms play a major role in diseases such as cancer. As a result, insights into the molecular details are needed to advance our understanding, improve biological models, and develop novel therapeutics. Dynamic single-molecule technologies offer an exciting opportunity to meet these challenges and to study protein function and activity in real time. Here, we present our efforts for enabling discoveries in the field of DNA repair using the combination of fluorescence microscopy with optical tweezers. This technology has been used to study CMG helicase dynamics, the role of BRCA2 in RAD51 nucleation, and the mechanism of the DNA repair complex FANCD2-FANCI. It is also suitable for many applications involving mismatch repair (MMR), base excision repair (BER), and nucleotide excision repair (NER), among other lesions. We present several examples in which our technologies and biochemistry enhanced the understanding of these systems. Importantly, we show that advances in dynamic single-molecule methods can be turned into an easy-to-use and stable instrument that is unlocking new avenues for studying DNA-binding proteins in the field of DNA repair. Citation Format: Paul Pease, Jack O'Sullivan, Trey Simpson, Josephine Yeh, Andrea Candelli. Unlocking DNA repair mechanisms with dynamic single-molecule analysis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 1480.