Department of Genome Sciences, University of Washington, Seattle, Washington, USA.
Department of Pathology, University of Washington, Seattle, Washington, USA.
Nat Methods. 2015 Jan;12(1):71-8. doi: 10.1038/nmeth.3205. Epub 2014 Dec 1.
High-throughput methods based on chromosome conformation capture have greatly advanced our understanding of the three-dimensional (3D) organization of genomes but are limited in resolution by their reliance on restriction enzymes. Here we describe a method called DNase Hi-C for comprehensively mapping global chromatin contacts. DNase Hi-C uses DNase I for chromatin fragmentation, leading to greatly improved efficiency and resolution over that of Hi-C. Coupling this method with DNA-capture technology provides a high-throughput approach for targeted mapping of fine-scale chromatin architecture. We applied targeted DNase Hi-C to characterize the 3D organization of 998 large intergenic noncoding RNA (lincRNA) promoters in two human cell lines. Our results revealed that expression of lincRNAs is tightly controlled by complex mechanisms involving both super-enhancers and the Polycomb repressive complex. Our results provide the first glimpse of the cell type-specific 3D organization of lincRNA genes.
基于染色体构象捕获的高通量方法极大地提高了我们对基因组三维(3D)结构的理解,但由于依赖于限制性内切酶,其分辨率有限。在这里,我们描述了一种称为 DNase Hi-C 的方法,用于全面绘制全基因组染色质接触图谱。DNase Hi-C 使用 DNase I 进行染色质片段化,与 Hi-C 相比,其效率和分辨率有了显著提高。将这种方法与 DNA 捕获技术相结合,为精细染色质结构的靶向作图提供了一种高通量方法。我们应用靶向 DNase Hi-C 来描述两种人类细胞系中 998 个大型长间隔非编码 RNA(lincRNA)启动子的 3D 结构。我们的结果表明,lincRNA 的表达受到涉及超级增强子和多梳抑制复合物的复杂机制的严格控制。我们的结果首次揭示了 lincRNA 基因的细胞类型特异性 3D 组织。