Takanashi Kenta, Kato Teru
Graduate School of Bionics, Computer and Media Sciences, Tokyo University of Technology, 1404-1 Katakura, Hachioji, Tokyo 192-0982, Japan.
Analyst. 2014 May 7;139(9):2122-6. doi: 10.1039/c4an00016a.
DNA methylation is an epigenetic mechanism for transcriptional regulation. The methylation process controls cellular differentiation and is defective in many diseases including cancer. Therefore, the development of a simple method for analysing cytosine methylation in a target gene is required. Here we report a conceptually new method for sequence-selective chemical modification of a single cytosine in single-stranded DNA (ssDNA) using two DNA probes to form a DNA three-way junction with the ssDNA. The method was successfully used in a simple quantitative polymerase-chain-reaction-based assay for discrimination of a single methylated cytosine.
DNA甲基化是一种用于转录调控的表观遗传机制。甲基化过程控制细胞分化，并且在包括癌症在内的许多疾病中存在缺陷。因此，需要开发一种用于分析目标基因中胞嘧啶甲基化的简单方法。在此，我们报告了一种概念上全新的方法，该方法利用两个DNA探针与单链DNA（ssDNA）形成DNA三链体连接，对单链DNA中的单个胞嘧啶进行序列选择性化学修饰。该方法已成功应用于一种基于简单定量聚合酶链反应的检测中，用于鉴别单个甲基化胞嘧啶。
