Oldenburg Anja R, Collas Philippe
Department of Molecular Medicine, Institute of Basic Medical Sciences, University of Oslo, Blindern, 0317, 1112, Oslo, Norway.
Methods Mol Biol. 2016;1411:315-24. doi: 10.1007/978-1-4939-3530-7_20.
The nuclear lamina is a meshwork of A- and B-type lamins which interact with chromatin and regulate many nuclear functions. Recent studies have reported the discovery of chromatin domains interacting with nuclear lamins by chromatin immunoprecipitation (ChIP) of lamin A/C or B1 and identification of the associated DNA sequences by microarray or high-throughput sequencing. ChIP has been used over many years to get a snapshot of interactions between DNA and proteins in cells, including modified histones, transcription factors, chromatin remodelers, and recently, structural proteins such as nuclear lamins. We describe here the procedure we have established in our laboratory for ChIP of lamin A/C and lamin B1 from human cultured cells. The protocol is compatible with polymerase chain reaction and high-throughput sequencing analysis of the co-immunoprecipitated DNA.
核纤层是由 A 型和 B 型核纤层蛋白组成的网络结构，它与染色质相互作用并调节许多核功能。最近的研究报道了通过对核纤层蛋白 A/C 或 B1 进行染色质免疫沉淀（ChIP）发现与核纤层相互作用的染色质结构域，并通过微阵列或高通量测序鉴定相关的 DNA 序列。多年来，ChIP 一直被用于获取细胞中 DNA 与蛋白质之间相互作用的快照，包括修饰的组蛋白、转录因子、染色质重塑因子，以及最近的核纤层等结构蛋白。我们在此描述了我们实验室建立的从人培养细胞中对核纤层蛋白 A/C 和核纤层蛋白 B1 进行 ChIP 的方法。该方案与共免疫沉淀 DNA 的聚合酶链反应和高通量测序分析兼容。
Zotero 插件