Department of Biological & Biomedical Sciences, Rowan University, Glassboro, NJ, USA.
Methods Mol Biol. 2025;2866:99-109. doi: 10.1007/978-1-0716-4192-7_6.
Identification of the occupancy of transcription factors (TFs) and nucleosomes across the genome yields insights into the regulation of gene expression patterns. While several independent techniques can be performed and then analyzed in composite to reveal this chromatin landscape, the use of micrococcal nuclease (MNase) digestion can resolve the footprints of nearly all chromatin proteins simultaneously. The protocol below describes the use of MNase to identify chromatin footprints of both TFs and nucleosomes in two vastly different cell types, Mouse embryonic stem cells (mESCs) and sperm, with differing levels of chromatin compaction.
鉴定转录因子 (TFs) 和核小体在整个基因组中的占据情况,可以深入了解基因表达模式的调控。虽然可以同时使用几种独立的技术进行分析,然后综合分析来揭示这种染色质图谱,但使用微球菌核酸酶 (MNase) 消化可以同时解析几乎所有染色质蛋白的足迹。下面的方案描述了使用 MNase 来鉴定两种差异很大的细胞类型(小鼠胚胎干细胞 (mESCs) 和精子)中 TF 和核小体的染色质足迹,这两种细胞类型的染色质紧凑程度不同。