Cancer Epigenetics Group, Josep Carreras Leukaemia Research Institute (IJC), Barcelona, Spain.
Life Sciences Department, Barcelona Supercomputing Center (BSC), Barcelona, Spain.
Epigenetics. 2022 Dec;17(12):1677-1685. doi: 10.1080/15592294.2022.2053816. Epub 2022 Mar 21.
Mouse has been extensively used as a model organism in many studies to characterize biological pathways and drug effects and to mimic human diseases. Similar DNA sequences between both species facilitate these types of experiments. However, much less is known about the mouse epigenome, particularly for DNA methylation. Progress in delivering mouse DNA methylomes has been slow due to the currently available time-consuming and expensive methodologies. Following the great acceptance of the human DNA methylation microarrays, we have herein validated a newly developed DNA methylation microarray (Infinium Mouse Methylation BeadChip) that interrogates 280,754 unique CpG sites within the mouse genome. The CpGs included in the platform cover CpG Islands, shores, shelves and open sea sequences, and loci surrounding transcription start sites and gene bodies. From a functional standpoint, mouse ENCODE representative DNase hypersensitivity sites (rDHSs) and candidate cis-Regulatory Elements (cCREs) are also included. Herein, we show that the profiled mouse DNA methylation microarray provides reliable values among technical replicates; matched results from fresh frozen versus formalin-fixed samples; detects hemimethylated X-chromosome and imprinted CpG sites; and is able to determine CpG methylation changes in mouse cell lines treated with a DNA demethylating agent or upon genetic disruption of a DNA methyltransferase. Most important, using unsupervised hierarchical clustering and t-SNE approaches, the platform is able to classify all types of normal mouse tissues and organs. These data underscore the great features of the assessed microarray to obtain comprehensive DNA methylation profiles of the mouse genome.
小鼠已被广泛用作许多研究中的模式生物,以研究生物途径和药物作用,并模拟人类疾病。两种物种之间相似的 DNA 序列有助于进行此类实验。然而,人们对小鼠的表观基因组知之甚少,特别是对 DNA 甲基化。由于目前可用的耗时且昂贵的方法学,提供小鼠 DNA 甲基组学的进展一直很缓慢。在人类 DNA 甲基化微阵列得到广泛接受之后,我们在此验证了一种新开发的 DNA 甲基化微阵列(Infinium Mouse Methylation BeadChip),该微阵列可检测小鼠基因组内 280754 个独特的 CpG 位点。该平台中包含的 CpG 覆盖了 CpG 岛、海岸、支架和开放海域序列,以及转录起始位点和基因体周围的基因座。从功能角度来看,还包括小鼠 ENCODE 代表性 DNase 超敏位点(rDHS)和候选顺式调控元件(cCRE)。在此,我们表明,所分析的小鼠 DNA 甲基化微阵列在技术重复中提供了可靠的值;新鲜冷冻与福尔马林固定样本的结果匹配;检测到半甲基化 X 染色体和印迹 CpG 位点;并能够确定用 DNA 去甲基化剂处理的小鼠细胞系或 DNA 甲基转移酶遗传缺失后的 CpG 甲基化变化。最重要的是,使用无监督层次聚类和 t-SNE 方法,该平台能够对所有类型的正常小鼠组织和器官进行分类。这些数据突出了评估的微阵列的出色功能,可获得小鼠基因组的全面 DNA 甲基化图谱。