One-pot approach for examining the DNA methylation patterns using an engineered methyl-probe
One-pot approach for examining the DNA methylation patterns using an engineered methyl-probe
复制标题
DOI：
10.1016/j.bios.2014.02.064
复制
发表时间：
2014-08-15
影响因子：
12.6
通讯作者：
Yuan, Chongli
中科院分区：
文献类型：
--
作者：
Kim, Seong-Eun;Chang, Matthew;Yuan, Chongli
异常的DNA甲基化是在各种类型的人类癌症中常见的观察结果，即，乳腺癌和肺癌。然而，目前的DNA甲基化检测方法需要亚硫酸氢盐处理，并且进行起来费力或昂贵。为了应对这些挑战，我们开发了一种基于MBD 1蛋白的甲基探针。结合荧光相关光谱，我们的探针可以灵敏地检测DNA甲基化的存在，在一锅法测定浓度高于20 nM。该探针可以定量高于20 nM的甲基化CG二核苷酸的总量，与DNA序列背景、浓度（20-1900 nM）和甲基化水平（5-100%）无关。我们的检测平台提供了一种简单而廉价的替代DNA甲基化检测方法。(c)2014爱思唯尔有限公司版权所有。
Aberrant DNA methylation is a common observation in various types of human cancers, i.e., breast and lung cancers. Nevertheless, the current DNA methylation detection approaches require bisulfite treatments and are laborious or costly to perform. To address these challenges, we developed a methyl-probe based on the MBD1 protein. Combined with fluorescence correlation spectroscopy, our probe can sensitively detect the existence of DNA methylation at concentrations above 20 nM in a one-pot assay. The probe can quantify the total amount of methylated CG dinucleotides above similar to 20 nM, independent of DNA sequence contexts, concentrations (20-1900 nM) and methylation levels (5-100%). Our detection platform offers a simple and cheap alternative DNA methylation detection approach. (c) 2014 Elsevier B.V. All rights reserved.