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

总结 蛋白质-DNA相互作用是DNA依赖性过程的基础，例如转录，复制， DNA修复和染色体组织在细胞生命的所有领域。基于富集的检测方法 蛋白质-DNA相互作用被广泛用于获得转录因子的全基因组高分辨率图谱 结合和染色质状态。然而，这些方法在应用于单细胞时具有严重的缺点。 在单细胞应用中，基于富集的方法不能区分未结合的DNA和未结合的DNA。 蛋白质结合DNA的不成功回收，因为两者都导致DNA片段缺失和数据稀疏 这使得难以或不可能获得起始于转录因子的细胞类型分辨的结合谱， 复杂的组织这种限制大大减少了单细胞数据的信息内容。MACHA将 检测细胞中蛋白质-DNA相互作用，如由外源GpC催化的胞嘧啶甲基化 甲基转移酶（GpCMTase），其通过识别甲基转移酶（GpCMTase）的第一抗体导向蛋白质结合的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.