Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore.
Methods. 2012 Nov;58(3):289-99. doi: 10.1016/j.ymeth.2012.08.009. Epub 2012 Aug 25.
Long-range chromatin contacts between specific DNA regulatory elements play a pivotal role in gene expression regulation, and a global characterization of these interactions in the 3-dimensional (3D) chromatin structure is imperative in understanding signaling networks and cell states. Chromatin Interaction Analysis using Paired-End Tag sequencing (ChIA-PET) is a method which converts functional chromatin structure into millions of short tag sequences. Combining Chromatin Immunoprecipitation (ChIP), proximity ligation and high-throughput sequencing, ChIA-PET provides a global and unbiased interrogation of higher-order chromatin structures associated with specific protein factors. Here, we describe the detailed procedures of the ChIA-PET methodology, unraveling transcription-associated chromatin contacts in a model human cell line.
长程染色质相互作用在特定 DNA 调控元件之间发挥着关键作用,对这些相互作用在三维染色质结构中的全面描述对于理解信号网络和细胞状态至关重要。使用配对末端标签测序(ChIA-PET)进行染色质相互作用分析是一种将功能染色质结构转化为数百万个短标签序列的方法。结合染色质免疫沉淀(ChIP)、邻近连接和高通量测序,ChIA-PET 提供了对与特定蛋白因子相关的高级染色质结构的全面、无偏检测。在这里,我们描述了 ChIA-PET 方法学的详细步骤,揭示了模型人类细胞系中与转录相关的染色质接触。