10-11 易位 (TET) 蛋白通过将 5-甲基胞嘧啶氧化为 5-羟甲基胞嘧啶 (5hmC) 来协调脱氧核糖核酸 (DNA) 甲基化-去甲基化动力学，并且在各种癌症中经常失活。由于 5hmC 作为表观遗传生物标志物对于癌症诊断、发病机制和治疗的重要性，因此对其快速、精确的定量至关重要。在这里，我们报告了一种高度灵敏的电化学方法，使用石墨烯片定量基因组 5hmC，该石墨烯片通过单步电学方法用生物素和金纳米颗粒 (Bt-AuNP) 进行电化学剥离和功能化。 Bt-AuNPs 与石墨烯的结合增强了含有 5hmC 的 DNA 的特异性，并增强了 DNA 中 5hmC 氧化为 5-甲酰胞嘧啶的能力。当与金电极耦合时，基于 Bt-AuNP-石墨烯的传感器在检测 5hmC 方面表现出卓越的灵敏度和特异性，检测限为 63.2 fM。此外，我们的传感器表现出卓越的能力，可以测量一系列生物样品的 5hmC 水平，包括由于 TET 基因破坏或致癌转化而具有不同 5hmC 水平的临床前小鼠组织，以及人类前列腺癌细胞系。因此，我们的传感策略在癌症诊断和预后方面具有巨大潜力。© 2024 作者。由爱思唯尔有限公司出版

Ten-eleven translocation (TET) proteins orchestrate deoxyribonucleic acid (DNA) methylation-demethylation dynamics by oxidizing 5-methylcytosine to 5-hydroxymethylcytosine (5hmC) and are frequently inactivated in various cancers. Due to the significance of 5hmC as an epigenetic biomarker for cancer diagnosis, pathogenesis, and treatment, its rapid and precise quantification is essential. Here, we report a highly sensitive electrochemical method for quantifying genomic 5hmC using graphene sheets that were electrochemically exfoliated and functionalized with biotin and gold nanoparticles (Bt-AuNPs) through a single-step electrical method. The attachment of Bt-AuNPs to graphene enhances the specificity of 5hmC-containing DNA and augments the oxidation of 5hmC to 5-formylcytosine in DNA. When coupled to a gold electrode, the Bt-AuNP-graphene-based sensor exhibits exceptional sensitivity and specificity for detecting 5hmC, with a detection limit of 63.2 fM. Furthermore, our sensor exhibits a remarkable capacity to measure 5hmC levels across a range of biological samples, including preclinical mouse tissues with varying 5hmC levels due to either TET gene disruption or oncogenic transformation, as well as human prostate cancer cell lines. Therefore, our sensing strategy has substantial potential for cancer diagnostics and prognosis.© 2024 The Authors. Published by Elsevier Ltd.