Raurell-Vila Helena, Ramos-Rodríguez Mireia, Pasquali Lorenzo
Program of Predictive and Personalized Medicine of Cancer (PMPPC), Endocrine Regulatory Genomics Laboratory, Department of Endocrinology and Nutrition, Germans Trias i Pujol University Hospital and Research Institute, Badalona, Spain.
Josep Carreras Leukaemia Research Institute, Badalona, Spain.
Methods Mol Biol. 2018;1766:197-208. doi: 10.1007/978-1-4939-7768-0_11.
The regulatory mechanisms that ensure an accurate control of gene transcription are central to cellular function, development and disease. Such mechanisms rely largely on noncoding regulatory sequences that allow the establishment and maintenance of cell identity and tissue-specific cellular functions.The study of chromatin structure and nucleosome positioning allowed revealing transcription factor accessible genomic sites with regulatory potential, facilitating the comprehension of tissue-specific cis-regulatory networks. Recently a new technique coupled with high-throughput sequencing named Assay for Transposase Accessible Chromatin (ATAC-seq) emerged as an efficient method to chart open chromatin genome wide. The application of such technique to different cell types allowed unmasking tissue-specific regulatory elements and characterizing cis-regulatory networks. Herein we describe the implementation of the ATAC-seq method to human pancreatic islets, a tissue playing a central role in the control of glucose metabolism.
确保基因转录精确控制的调控机制是细胞功能、发育和疾病的核心。这些机制很大程度上依赖于非编码调控序列,这些序列允许建立和维持细胞身份以及组织特异性细胞功能。对染色质结构和核小体定位的研究有助于揭示具有调控潜力的转录因子可及基因组位点,促进对组织特异性顺式调控网络的理解。最近,一种与高通量测序相结合的新技术——转座酶可及染色质分析(ATAC-seq),成为一种在全基因组范围内绘制开放染色质的有效方法。将这种技术应用于不同细胞类型,有助于揭示组织特异性调控元件并表征顺式调控网络。在此,我们描述了将ATAC-seq方法应用于人类胰岛的过程,胰岛是在葡萄糖代谢控制中起核心作用的组织。