Roberti Annalisa, Fraga Mario Fernández
Cancer Epigenetics and Nanomedicine Laboratory, Nanomaterials and Nanotechnology Research Center (CINN), Spanish National Research Council (CSIC), El Entrego, Spain.
Instituto de Investigación Sanitaria del Principado de Asturias (ISPA), Oviedo, Spain.
Methods Mol Biol. 2025;2930:229-243. doi: 10.1007/978-1-0716-4558-1_16.
Alterations in the epigenetic landscape are now recognized as a hallmark of cancer. Among these, histone posttranslational modifications (HPTMs) represent a critical control layer, playing a significant role in chromatin remodeling and gene expression regulation. The comprehensive mapping of these modifications on a genome-wide scale is crucial for identifying regulatory regions and understanding downstream pathways, thereby providing a deeper and more integrated understanding of cancer biology. CUT&RUN (Cleavage Under Targets and Release Using Nuclease), combined with next-generation sequencing, is emerging as a highly reliable, low-background, and user-friendly technology for mapping antibody-bound chromatin regions in a genome-wide manner. In this chapter, we provide a comprehensive overview of an optimized protocol for targeting HPTMs in frozen tissue samples, emphasizing the crucial role of nucleus purification in our methodology.
表观遗传格局的改变现已被公认为癌症的一个标志。其中，组蛋白翻译后修饰（HPTMs）代表了一个关键的控制层，在染色质重塑和基因表达调控中发挥着重要作用。在全基因组范围内对这些修饰进行全面定位对于识别调控区域和理解下游通路至关重要，从而能更深入、更全面地理解癌症生物学。CUT&RUN（靶向切割并利用核酸酶释放）与新一代测序相结合，正成为一种高度可靠、背景低且用户友好的技术，可在全基因组范围内绘制与抗体结合的染色质区域。在本章中，我们全面概述了一种针对冷冻组织样本中HPTMs的优化方案，强调了细胞核纯化在我们方法中的关键作用。
