Direct single nucleotide mapping of genomic CpG marks

描述(由申请人提供)： 在过去的十年里，表观遗传现象声称在细胞调控过程中发挥了核心作用，并被证明是理解复杂的人类疾病的重要因素。最被理解的表观遗传机制之一是DNA甲基化。在哺乳动物基因组中，胞嘧啶(C)存在于两种功能状态：未甲基化或嘧啶环5位甲基化(5mC)。最近对人类和小鼠脑、神经元和小鼠胚胎干细胞基因组DNA的研究发现，CpG二核苷酸中相当大一部分5mC在加氧酶的作用下转化为5-羟甲基胞嘧啶(HMC)。这些发现为长期难以捉摸的活性DNA去甲基化机制提供了重要线索，并支持了哺乳动物表观遗传调控领域的新一轮研究。然而，现有的全基因组表观基因组分析实验技术的缺陷阻碍了这类研究。该项目致力于开发一种突破性技术，用于全基因组分析CpG二核苷酸的修饰状态，将单碱基分辨率与基因组的定向和经济测序相结合。

DESCRIPTION (provided by applicant): Over the past decade, epigenetic phenomena claimed a central role in cell regulatory processes and proved important factors for understanding complex human diseases. One of the best understood epigenetic mechanisms is DNA methylation. In the mammalian genome, cytosines (C) were long known to exist in two functional states: unmethylated or methylated at the 5-position of the pyrimidine ring (5mC). Recent studies of genomic DNA from the human and mouse brain, neurons and from mouse embryonic stem cells found that a substantial fraction of 5mC in CpG dinucleotides is converted to 5-hydroxymethylcytosine (hmC) by the action of oxygenases. These findings provided important clues in a long elusive mechanism of active DNA demethylation and bolstered a fresh wave of studies in the area of epigenetic regulation in mammals. However, such studies are hindered by the shortcomings of available experimental techniques for genome-wide epigenome analysis. This project is dedicated to developing a break-through technique for genome-wide analysis of the modification status of CpG dinucleotides that combines single-base resolution with targeted and economic sequencing of the genome.