Single Molecule Epigenetic Analysis in a Nanofluidic Channel
Single Molecule Epigenetic Analysis in a Nanofluidic Channel
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DOI：
10.1021/ac9028642
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发表时间：
2010-03-15
影响因子：
7.4
通讯作者：
Soloway, Paul D.
中科院分区：
文献类型：
--
作者：
Cipriany, Benjamin R.;Zhao, Ruqian;Soloway, Paul D.
表观遗传状态受DNA甲基化和与DNA结合的组蛋白的大量修饰控制。这些状态对于适当的发育调节基因表达是必不可少的，并且在许多疾病中受到干扰。人们对鉴定全基因组范围内的表观遗传标记位置以及了解这些标记在细胞类型之间如何随着环境的变化或根据健康和疾病状态而变化非常感兴趣。目前的表观基因组分析采用亚硫酸氢盐测序和染色质免疫沉淀，但一次只能查询一种类型的表观遗传标记，DNA甲基化或组蛋白修饰，并且通常需要大量的输入材料。为了克服这些局限性，我们建立了一种方法，使用纳米流体和荧光显微镜检测DNA和组蛋白在个别染色质片段在约10 Mbp/min。我们证明了其实用性的表观遗传分析，通过识别DNA甲基化的个别分子。该技术将为全基因组范围内同时分析单个分子上的多种表观遗传状态提供前所未有的机会。
Epigenetic states are governed by DNA methylation and a host of modifications to histones bound with DNA. These states are essential for proper developmentally regulated gene expression and are perturbed in many diseases. There is great interest in identifying epigenetic mark placement genome wide and understanding how these marks vary among cell types, with changes in environment or according to health and disease status. Current epigenomic analyses employ bisulfite sequencing and chromatin immunoprecipitation, but query only one type of epigenetic mark at a time, DNA methylation, or histone modifications and often require substantial input material. To overcome these limitations, we established a method using nanofluidics and multicolor fluorescence microscopy to detect DNA and histones in individual chromatin fragments at about 10 Mbp/min. We demonstrated its utility for epigenetic analysis by identifying DNA methylation on individual molecules. This technique will provide the unprecedented opportunity for genome wide, simultaneous analysis of multiple epigenetic states on single molecules.