New insights into the cis-regulation of the CFTR gene in pancreatic cells [CUT&Run]

To understand the role of 3D chromatin architecture in establishing tissue-specific expression of the CFTR gene, we mapped chromatin interactions using Circular Chromosome Conformation Capture (4C) and epigenomic data such as H3K27ac and Assay for Transposase-Accessible Chromatin (ATAC-seq) on Capan-1 pancreatic cells. Overall design: We used CUT&RUN with H3K27ac to identify active enhancer regions in Capan-1 cells. We also mapped CTCF, H3K27ac and H3K27me3 binding site in Caco-2 cells.

为阐明3D染色质构象在调控CFTR基因（囊性纤维化跨膜传导调节因子基因）组织特异性表达中的作用，我们以Capan-1胰腺细胞为研究模型，通过环状染色质构象捕获（4C，Circular Chromosome Conformation Capture）技术绘制染色质相互作用图谱，并结合表观基因组学数据，包括组蛋白H3第27位赖氨酸乙酰化（H3K27ac）与转座酶可及性染色质测序（ATAC-seq，Assay for Transposase-Accessible Chromatin）数据。整体实验设计如下：我们采用靶标切割与释放技术（CUT&RUN）结合H3K27ac标记，以鉴定Capan-1细胞中的活性增强子区域；同时还在Caco-2细胞中绘制了CCCTC结合因子（CTCF）、H3K27ac及组蛋白H3第27位赖氨酸三甲基化（H3K27me3）的结合位点图谱。