• Electrochemically Detecting DNA Methylation in the EN1 gene Promoter: Implications for understanding Ageing and Disease

      Morgan, Amy; Acutt, Katie; Mc Auley, Mark; University of Chester
      There is a growing need for biomarkers which predict age-onset pathology. Although this is challenging, the methylome offers significant potential. Cancer is associated with the hypermethylation of many gene promoters, among which are developmental genes. Evolutionary theory suggests developmental genes arbitrate early-late life trade-offs, causing epimutations that increase disease vulnerability. Such genes could predict age related disease. The aim of this work was to optimise an electrochemical procedure for the future investigation of a broad range of ageing related pathologies. An electrochemical approach, which adopted three analytical techniques, was used to investigate DNA methylation in the EN1 gene promoter. Using synthetic single stranded DNA, one technique was able to detect DNA at concentrations as low as 10nM, with methylation status distinguishable at concentrations >25nM. A negative correlation could be observed between % methylation of heterogeneous solution and the key electrochemical parameter, Rct (r = -0.982, p < 0.01). The technique was applied to the breast cancer cell line MCF-7, where a similar correlation was observed (r = -0.965, p < 0.01). These results suggest electrochemistry can effectively measure DNA methylation at low concentrations of DNA. This has implications for the future detection of age-related disease.