Abstract：
Histone post-translational modifications (PTMs) influence the overall structure of the chromatin and gene expression. Over the course of cell differentiation, the distribution of histone modifications is remodeled, resulting in cell type-specific patterns. In the past, their study was limited to abundant cell types that could be purified in necessary numbers. However, studying these cell type-specific dynamic changes in heterogeneous in vivo settings requires sensitive single-cell methods. Current advances in single-cell sequencing methods remove these limitations, allowing the study of nonpurifiable cell types. One complicating factor is that some of the most biologically interesting cell types, including stem and progenitor cells that undergo differentiation, only make up a small fraction of cells in a tissue. This makes whole-tissue analysis rather inefficient. In this chapter, we present a sort-assisted single-cell Chromatin ImmunoCleavage sequencing technique (sortChIC) to map histone PTMs in single cells. This technique combines the mapping of histone PTM location in combination with surface staining-based enrichment, to allow the integration of established strategies for rare cell type enrichment. In general terms, this will enable researchers to quantify local and global chromatin changes in dynamic complex biological systems and can provide additional information on their contribution to lineage and cell-type specification in physiological conditions and disease.
摘要：
组蛋白翻译后修饰(PTM)影响染色质的整体结构和基因表达。在细胞分化过程中，组蛋白修饰的分布被重塑，导致细胞类型特定的模式。在过去,他们的研究仅限于可以纯化所需数量的丰富细胞类型。然而,在异质体内环境中研究这些细胞类型特异性的动态变化需要灵敏的单细胞方法.目前单细胞测序方法的进展消除了这些限制，允许研究不可纯化的细胞类型。一个复杂的因素是一些生物学上最有趣的细胞类型，包括经历分化的干细胞和祖细胞，只构成组织中的一小部分细胞。这使得整个组织分析相当低效。在这一章中,我们提出了一种排序辅助的单细胞染色质免疫裂解测序技术（sortChIC）来绘制单细胞中的组蛋白PTM。该技术将组蛋白PTM位置的映射与基于表面染色的富集相结合，以允许整合已建立的稀有细胞类型富集策略。总的来说,这将使研究人员能够量化动态复杂生物系统中的局部和全局染色质变化，并可以提供有关其在生理状况和疾病中对谱系和细胞类型规范的贡献的其他信息。
