通过甲基化 CpG 免疫沉淀法富集甲基化 DNA。

Enrichment of methylated DNA by methyl-CpG immunoprecipitation.

作者信息

Sonnet Miriam, Baer Constance, Rehli Michael, Weichenhan Dieter, Plass Christoph

机构信息

Department of Epigenomics and Cancer Risk Factors, German Cancer Research Center (DKFZ), Heidelberg, Germany.

出版信息

Methods Mol Biol. 2013;971:201-12. doi: 10.1007/978-1-62703-269-8_11.

DOI:10.1007/978-1-62703-269-8_11

PMID:23296965

Abstract

Normal DNA methylation is an epigenetic modification required for proper development. Aberrant DNA methylation, in contrast, is frequently observed in many different malignancies including leukemias and lymphomas. Global DNA methylation profiling addresses the methylated sequences (methylome) of patient genomes to identify disease-specific methylation patterns. Workload in methylome analyses can be considerably reduced by methylome enrichment using proteins or antibodies with high affinity to methylated DNA. Methyl-CpG Immunoprecipitation (MCIp) employs an immobilized recombinant human methyl-CpG binding domain protein 2, MBD2, which binds methylated CpGs in double-stranded DNA. Elution with increasing salt concentrations allows the fractionated enrichment of different degrees of methylation.

摘要

正常的DNA甲基化是正常发育所需的一种表观遗传修饰。相比之下，异常的DNA甲基化在包括白血病和淋巴瘤在内的许多不同恶性肿瘤中经常被观察到。全基因组DNA甲基化分析旨在研究患者基因组的甲基化序列（甲基化组），以识别疾病特异性的甲基化模式。通过使用对甲基化DNA具有高亲和力的蛋白质或抗体进行甲基化组富集，可以显著减少甲基化组分析的工作量。甲基化CpG免疫沉淀（MCIp）使用固定化的重组人甲基化CpG结合域蛋白2（MBD2），该蛋白可结合双链DNA中的甲基化CpG。用逐渐增加的盐浓度进行洗脱，可以对不同程度的甲基化进行分级富集。