Department of Molecular Biology, Princeton University, Princeton, NJ, USA.
Methods Mol Biol. 2024;2805:127-135. doi: 10.1007/978-1-0716-3854-5_8.
The modulation of cis-regulatory elements (e.g., enhancers and promoters) is a major mechanism by which gene expression can be controlled in a temporal and spatially restricted manner. However, methods for both identifying these elements and inferring their activity are limited and often require a substantial investment of time, money, and resources. Here, using mammalian skin as a model, we demonstrate a streamlined protocol by which these hurdles can be overcome using a novel chromatin profiling technique (CUT&RUN) to map histone modifications genome-wide. This protocol can be used to map the location and activity of putative cis-regulatory elements, providing mechanistic insight into how differential gene expression is controlled in mammalian tissues.
顺式调控元件（例如增强子和启动子）的调节是基因表达在时空上受到限制的主要机制。然而，鉴定这些元件并推断其活性的方法是有限的，并且通常需要大量的时间、金钱和资源。在这里，我们使用哺乳动物皮肤作为模型，展示了一种简化的方案，该方案使用新型染色质分析技术（CUT&RUN）来绘制全基因组组蛋白修饰，从而克服了这些障碍。该方案可用于绘制假定顺式调控元件的位置和活性，为理解哺乳动物组织中差异基因表达如何受到控制提供了机制上的见解。
