Abstract：
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.
摘要：
靶标下的切割和使用核酸酶的释放(CUT&RUN)是检测DNA和DNA相关蛋白之间的特异性相互作用的方法。它对于表征全基因组转录因子或共调节因子的结合是有价值的。此外，它可以用于表观遗传分析，染色质可及性评估，并确定监管要素。与更常用的染色质免疫沉淀（ChIP）相比，CUT&RUN具有若干优点,包括原位方法以及不需要超声处理。然而,最大的优点是减少了CUT和RUN所需的细胞数量，这使得它对细胞数量有限的实验更具吸引力。在这一章中,我们描述了一种可靠的巨噬细胞CUT&RUN方案,该方案可以在2天内进行,并且包括文库制备,以便样品可以直接测序。
