State Key Laboratory of Bioelectronics, Southeast University, Nanjing, PR China.
Electrophoresis. 2010 Oct;31(21):3537-44. doi: 10.1002/elps.201000326.
DNA methylation is one of the most important epigenetic modification types, which plays a critical role in gene expression. High efficient surveying of whole genome DNA methylation has been aims of many researchers for long. Recently, the rapidly developed massively parallel DNA-sequencing technologies open the floodgates to vast volumes of sequence data, enabling a paradigm shift in profiling the whole genome methylation. Here, we describe a strategy, combining methylated DNA immunoprecipitation sequencing with peak search to identify methylated regions on a whole-genome scale. Massively parallel methylated DNA immunoprecipitation sequencing combined with methylation DNA immunoprecipitation was adopted to obtain methylated DNA sequence data from human leukemia cell line K562, and the methylated regions were identified by peak search based on Poison model. From our result, 140 958 non-overlapping methylated regions have been identified in the whole genome. Also, the credibility of result has been proved by its strong correlation with bisulfite-sequencing data (Pearson R(2)=0.92). It suggests that this method provides a reliable and high-throughput strategy for whole genome methylation identification.
DNA 甲基化是最重要的表观遗传修饰类型之一,在基因表达中起着关键作用。长期以来,高效地检测全基因组 DNA 甲基化一直是许多研究人员的目标。最近,快速发展的大规模平行 DNA 测序技术为大量的序列数据打开了大门,使全基因组甲基化的分析发生了范式转变。在这里,我们描述了一种策略,将甲基化 DNA 免疫沉淀测序与峰搜索相结合,以在全基因组范围内识别甲基化区域。采用大规模平行的甲基化 DNA 免疫沉淀测序与甲基化 DNA 免疫沉淀相结合的方法,从人白血病细胞系 K562 中获得甲基化 DNA 序列数据,并基于泊松模型通过峰搜索来识别甲基化区域。在全基因组范围内,共鉴定到 140958 个非重叠的甲基化区域。此外,该结果与亚硫酸氢盐测序数据(Pearson R²=0.92)具有很强的相关性,证明了结果的可信度。这表明该方法为全基因组甲基化鉴定提供了一种可靠的高通量策略。