Institute of Functional Epigenetics (IFE), Helmholtz Zentrum München, Neuherberg, Germany.
Faculty of Biology, Ludwig-Maximilians-Universität München, Planegg-Martinsried, Germany.
EMBO Rep. 2024 Aug;25(8):3202-3220. doi: 10.1038/s44319-024-00199-2. Epub 2024 Aug 2.
In eukaryotes, DNA is packaged into chromatin with the help of highly conserved histone proteins. Together with DNA-binding proteins, posttranslational modifications (PTMs) on these histones play crucial roles in regulating genome function, cell fate determination, inheritance of acquired traits, cellular states, and diseases. While most studies have focused on individual DNA-binding proteins, chromatin proteins, or histone PTMs in bulk cell populations, such chromatin features co-occur and potentially act cooperatively to accomplish specific functions in a given cell. This review discusses state-of-the-art techniques for the simultaneous profiling of multiple chromatin features in low-input samples and single cells, focusing on histone PTMs, DNA-binding, and chromatin proteins. We cover the origins of the currently available toolkits, compare and contrast their characteristic features, and discuss challenges and perspectives for future applications. Studying the co-occurrence of histone PTMs, DNA-binding proteins, and chromatin proteins in single cells will be central for a better understanding of the biological relevance of combinatorial chromatin features, their impact on genomic output, and cellular heterogeneity.
在真核生物中,DNA 借助高度保守的组蛋白蛋白被包装成染色质。这些组蛋白上的翻译后修饰(PTMs)与 DNA 结合蛋白一起,在调节基因组功能、细胞命运决定、获得性状的遗传、细胞状态和疾病方面发挥着关键作用。虽然大多数研究都集中在单个 DNA 结合蛋白、染色质蛋白或组蛋白 PTMs 的批量细胞群体上,但这些染色质特征共同发生,并可能协同作用,在特定细胞中完成特定功能。这篇综述讨论了同时分析低输入样本和单细胞中多种染色质特征的最新技术,重点介绍了组蛋白 PTMs、DNA 结合和染色质蛋白。我们介绍了目前可用工具包的起源,比较和对比了它们的特征,并讨论了未来应用的挑战和前景。研究单个细胞中组蛋白 PTMs、DNA 结合蛋白和染色质蛋白的共同发生,将是更好地理解组合染色质特征的生物学相关性、它们对基因组输出和细胞异质性的影响的核心。