SAFU, Department of Research, Advanced Diagnostic and Technological Innovation, IRCCS Regina Elena National Cancer Institute, Rome, Italy.
Methods Mol Biol. 2022;2535:171-185. doi: 10.1007/978-1-0716-2513-2_14.
The transcription of each gene is tightly regulated by elements like promoters, enhancers, silencers and insulators. These elements determine the temporal and tissue-specific expression in development and disease. Drug resistance is the major obstacle in successfully treating cancer patients. In the recent years, it became evident that epigenetic changes represent one of the mechanisms that contribute to the onset and progression of cancer but also to the development of therapy resistance. The assay for transposase-accessible chromatin coupled with next generation sequencing (ATAC-seq) is a fast and easy technique to track epigenetic changes that result in different opening of the chromatin in regulatory regions genome-wide. The transposase cuts DNA in regions that are open and therefore accessible for transcription factors, regulatory RNAs and proteins that alter the architectural structure of the DNA and drive or inhibit transcription through the RNA polymerase. Here we describe a detailed protocol to perform an ATAC-seq of cells from culture or tissue.
每个基因的转录都受到启动子、增强子、沉默子和绝缘子等元件的严格调控。这些元件决定了在发育和疾病过程中的时空特异性表达。耐药性是成功治疗癌症患者的主要障碍。近年来,人们已经意识到,表观遗传变化是导致癌症发生和发展以及治疗耐药性发展的机制之一。转座酶可及染色质的检测与下一代测序(ATAC-seq)是一种快速而简单的技术,可跟踪导致染色质在调控区域内全基因组不同打开的表观遗传变化。转座酶在开放的区域切割 DNA,因此转录因子、调节 RNA 和蛋白质可以进入,这些因子和蛋白质可以改变 DNA 的结构,并通过 RNA 聚合酶驱动或抑制转录。在这里,我们描述了一个从培养细胞或组织中进行 ATAC-seq 的详细方案。