Department of Biological DNA Modification, Institute of Biotechnology, Vilnius University, Vilnius LT-02241, Lithuania.
Nat Commun. 2013;4:2190. doi: 10.1038/ncomms3190.
Dynamic patterns of cytosine-5 methylation and successive hydroxylation are part of epigenetic regulation in eukaryotes, including humans, which contributes to normal phenotypic variation and disease risk. Here we present an approach for the mapping of unmodified regions of the genome, which we call the unmethylome. Our technique is based on DNA methyltransferase-directed transfer of activated groups and covalent biotin tagging of unmodified CpG sites followed by affinity enrichment and interrogation on tiling microarrays or next generation sequencing. Control experiments and pilot studies of human genomic DNA from cultured cells and tissues demonstrate that, along with providing a unique cross-section through the chemical landscape of the epigenome, the methyltransferase-directed transfer of activated groups-based approach offers high precision and robustness as compared with existing affinity-based techniques.
动态的胞嘧啶-5 甲基化和连续羟化模式是真核生物（包括人类）表观遗传调控的一部分，有助于正常表型变异和疾病风险。在这里，我们提出了一种用于绘制基因组未修饰区域的方法，我们称之为未甲基化组。我们的技术基于 DNA 甲基转移酶定向转移活化基团和未修饰 CpG 位点的共价生物素标记，然后在平铺微阵列或下一代测序上进行亲和富集和检测。对照实验和来自培养细胞和组织的人类基因组 DNA 的初步研究表明，与现有的基于亲和性的技术相比，这种基于 DNA 甲基转移酶定向转移活化基团的方法不仅提供了对表观基因组化学景观的独特横断面，而且具有高精度和高稳健性。
