Single-cell and single-molecule profiling of protein-DNA interactions by MACHA

摘要 蛋白质-DNA相互作用是依赖DNA的过程的基础，例如转录、复制、 DNA修复，以及细胞生命各个领域的染色体组织。基于浓缩的方法来检测 蛋白质-DNA相互作用被广泛用于获得全基因组高分辨率转录因子图谱 结合状态和染色质状态。然而，当应用于单个细胞时，这些方法存在严重的缺陷。 在单细胞应用中，基于浓缩的方法不能区分未结合的DNA和 蛋白质结合的DNA回收不成功，因为两者都导致DNA片段缺失和数据稀疏 使得很难或不可能获得从 复杂的组织。这一限制大大减少了单元格数据的信息量。马查会 外源GPC催化胞嘧啶甲基化检测细胞中蛋白质与DNA的相互作用 甲基转移酶(GpCMTase)，由识别DNA的一抗直接作用于蛋白质结合的DNA。 感兴趣的蛋白质。

Summary Protein-DNA interactions are fundamental to DNA-dependent processes such as transcription, replication, DNA repair, and chromosome organization in all domains of cellular life. Enrichment-based methods to detect protein-DNA interactions are widely used to obtain genome-wide high-resolution maps of transcription factor binding and chromatin states. However, these methods have severe shortcomings when applied to single cells. In single-cell applications, enrichment-based methods cannot distinguish between unbound DNA and unsuccessful recovery of protein-bound DNA since both lead to absent DNA fragments and the sparse data makes it difficult or impossible to obtain cell-type-resolved binding profiles of transcription factors starting from complex tissues. This limitation significantly reduces the information content of single-cell data. MACHA will detect protein-DNA interactions in cells as cytosine methylation catalyzed by an exogenous GpC methyltransferase (GpCMTase) that is directed to protein-bound DNA by a primary antibody recognizing the protein of interest.