The_role_of_somatic_mutations_in_POLK_deficient_mouse_models__Duplex__

In this study, we propose to investigate the contribution of somatic mutations to DNA damage accumulated in NER-deficient mouse models. We will assess the mutation and indel burden in POLK-deficient mouse models and controls to understand the impact of translesion synthesis on the accumulation of somatic mutations. By investigating distinct tissue types, we will systematically analyse the tissue-specific accumulation of mutational signatures and DNA damage. This will include, but is not limited, to liver, kidney, lung and adrenal gland given their functional importance in disease. Frozen residual tissue will be received, paxgene-fixed and paraffin embedded and used for laser-capture microdissection (LCM) to dissect tissue individual histological locations in each tissue. In other cases, bulk DNA or tissue might be received. Extracted DNA from this procedure will be utilised for nanorate sequencing, a newly developed duplex sequencing protocol, will enable us to accurately detect somatic mutations as well as small insertions and deletions (indels). Overall, we anticipate the proposed study to improve our understanding of the role of somatic mutations in accelerated ageing phenotypes.