Metabolic Programming, TUM School of Life Sciences, Technical University of Munich, Freising, Germany.
Methods Mol Biol. 2024;2846:151-167. doi: 10.1007/978-1-0716-4071-5_10.
Cleavage Under Targets and Release Using Nuclease (CUT&RUN) is a method to detect specific interactions between DNA and DNA-associated proteins. It is valuable for the characterization of the binding of transcription factors or co-regulators genome wide. Furthermore, it can be used for epigenetic profiling, chromatin accessibility assessment, and identification of regulatory elements. Compared to the more commonly used chromatin immunoprecipitation (ChIP), CUT&RUN has several advantages including an in situ approach as well as no need for sonication. However, the biggest advantage is the reduced cell amounts that are required for CUT&RUN, which makes it more attractive for experiments with limited cell numbers. In this chapter, we describe a reliable CUT&RUN protocol for macrophages that can be performed within 2 days and includes a library preparation so that the sample can be directly sequenced.
靶向切割和释放核酸酶（CUT&RUN）是一种检测 DNA 与 DNA 相关蛋白之间特定相互作用的方法。它对于转录因子或共调节因子在全基因组范围内的结合特征具有重要价值。此外，它还可用于表观遗传分析、染色质可及性评估和调控元件的鉴定。与更常用的染色质免疫沉淀（ChIP）相比，CUT&RUN 具有几个优势，包括原位方法和不需要超声处理。然而，最大的优势是 CUT&RUN 需要的细胞数量减少，这使得它在细胞数量有限的实验中更具吸引力。在本章中，我们描述了一种适用于巨噬细胞的可靠 CUT&RUN 方案，该方案可在 2 天内完成，并包括文库制备，以便可以直接对样品进行测序。
