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Electrochemically Detecting DNA Methylation in the EN1 gene Promoter: Implications for understanding Ageing and Disease.
Bioscience Reports ( IF 3.8 ) Pub Date : 2020-11-02 , DOI: 10.1042/bsr20202571
Amy E. Morgan 1 , Katie D. Acutt 1 , Mark T. Mc Auley 1
Affiliation  

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.

中文翻译:

电化学检测EN1基因启动子中的DNA甲基化:对理解衰老和疾病的影响。

对预测年龄发病病理的生物标记物的需求不断增长。尽管这具有挑战性,但是甲基化组具有巨大的潜力。癌症与许多基因启动子的高甲基化有关,其中包括发育基因。进化论认为,发育基因决定了早期生命的权衡取舍,造成了增加疾病易感性的突变。这样的基因可以预测与年龄有关的疾病。这项工作的目的是优化一种电化学程序,以便将来对各种与衰老有关的病理学进行研究。电化学方法采用了三种分析技术,用于研究EN1基因启动子中的DNA甲基化。使用合成的单链DNA,一种技术能够检测低至10nM的DNA,在浓度> 25nM时甲基化状态明显。在异质溶液的甲基化百分比与关键的电化学参数Rct之间可观察到负相关(r = -0.982,p <0.01)。该技术应用于乳腺癌细胞系MCF-7,在其中观察到类似的相关性(r = -0.965,p <0.01)。这些结果表明,电化学可以有效地测量低浓度DNA时的DNA甲基化。这对将来发现与年龄有关的疾病有影响。p <0.01)。这些结果表明,电化学可以有效地测量低浓度DNA时的DNA甲基化。这对将来发现与年龄有关的疾病有影响。p <0.01)。这些结果表明,电化学可以有效地测量低浓度DNA时的DNA甲基化。这对将来发现与年龄有关的疾病有影响。
更新日期:2020-11-04
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