Defining the mutagenic impacts of DNA resection during double-strand break repair (360G-Wellcome-227014_Z_23_Z)

I will conduct in-depth investigations into the mutagenic impacts of resection around DNA double-strand breaks (DSBs). I will use a combined approach to first quantify the lengths of the different types of DSB resection and follow this with siRNA mediated depletion and chemical inhibition of all major resection enzymes to comprehensively elucidate the mechanisms of resection. Little is known about resection lengths at DSBs, and the overlapping roles of the long-range resection enzymes BLM, DNA2, EXO1 and EXD2 are poorly characterised. What determines recruitment of one long-range enzyme over another to a particular DSB and how the lengths of resection vary between them is unknown. Given that resection appears to be the major mutagenic process of DSB repair, a detailed understanding of these processes and their regulation is crucial for understanding DSB-induced mutagenesis. Finally, I will take various approaches to investigate the mutagenic impacts of these processes, identifying all mutations from rare point mutations to large-scale chromosome rearrangements, and crucially defining how these various enzymes are associated with this mutagenesis. The findings from this project will yield new insights into the mutagenic mechanisms of DSB-repair, significantly advancing our understanding of carcinogenesis and cancer progression, as well as other DSB-related diseases.