Distinguishing Active Versus Passive DNA Demethylation Using Illumina MethylationEPIC BeadChip Microarrays.
Distinguishing Active Versus Passive DNA Demethylation Using Illumina MethylationEPIC BeadChip Microarrays.
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DOI：
10.1007/978-1-0716-1294-1_7
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发表时间：
2021
期刊：
影响因子：
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通讯作者：
Rothbart SB
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文献类型：
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作者：
Tiedemann RL;Eden HE;Huang Z;Robertson KD;Rothbart SB
基因组DNA （CpG）中位于鸟嘌呤之前的胞嘧啶核苷酸的5碳位置是DNA甲基化（5mC）的常见靶点。DNA甲基化去除可以通过主动和被动两种机制发生。10 - 11易位酶（TETs）将5mC逐步氧化为5-羟甲基胞嘧啶（5hmC）、5-甲酰基胞嘧啶（5fC）和5-羧基胞嘧啶（5caC）。在没有DNA甲基化维持的情况下，5mC也可以通过连续的细胞分裂被动地去除。在本章中，我们描述了将et辅助亚硫酸氢盐（TAB）和氧化亚硫酸氢盐（OxBS）转化到Illumina MethylationEPIC BeadChIP （EPIC阵列）上的方法，并展示了如何使用这些技术来区分主动和被动DNA去甲基化。我们还描述了综合生物信息学管道，以促进这种分析。
The 5-carbon positions on cytosine nucleotides preceding guanines in genomic DNA (CpG) are common targets for DNA methylation (5mC). DNA methylation removal can occur through both active and passive mechanisms. Ten-eleven translocation enzymes (TETs) oxidize 5mC in a stepwise manner to 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine (5caC). 5mC can also be removed passively through sequential cell divisions in the absence of DNA methylation maintenance. In this chapter, we describe approaches that couple TET-assisted bisulfite (TAB) and oxidative bisulfite (OxBS) conversion to the Illumina MethylationEPIC BeadChIP (EPIC array) and show how these technologies can be used to distinguish active versus passive DNA demethylation. We also describe integrative bioinformatics pipelines to facilitate this analysis.