Simultaneous single-cell analysis of 5mC and 5hmC with SIMPLE-seq

We used a new technology, named SIMPLE-seq, to perform simultaneous profiling of 5mC and 5hmC on mESCs and PBMCs at single-cell,single molecule level. Overall design: We develop SIMPLE-seq for simultaneously profiling the genome-wide true methylome and hydroxymethylome quantitatively, in highly scalable single-cell manner. SIMPLE-seq measures both true 5mC and 5hmC on the same DNA molecule from one cell. To demonstrate the utility of SIMPLE-seq, we measured single-cell methylome and hydroxymethylome in mouse embryonic stem cells (mESC) and human peripheral blood mononuclear cells.本研究采用一种名为SIMPLE-seq的新技术，在单细胞、单分子水平上对小鼠胚胎干细胞（mouse embryonic stem cells, mESCs）和人外周血单个核细胞（peripheral blood mononuclear cells, PBMCs）的5-甲基胞嘧啶（5-methylcytosine, 5mC）与5-羟甲基胞嘧啶（5-hydroxymethylcytosine, 5hmC）开展同步谱分析。 整体实验设计：本研究开发的SIMPLE-seq技术，可实现高度可扩展的单细胞检测，能够对全基因组范围的真实甲基化组与羟甲基化组进行定量同步分析。该技术可对单个细胞来源的同一DNA分子上的真实5mC与5hmC同时完成检测。为验证SIMPLE-seq的应用价值，本研究针对小鼠胚胎干细胞（mESCs）及人外周血单个核细胞的单细胞甲基化组与羟甲基化组进行了检测。