通过生物素化的 dCas9 对特定基因座的 3D 染色质的空间构象和时间动态进行多重捕获。
Multiplexed capture of spatial configuration and temporal dynamics of locus-specific 3D chromatin by biotinylated dCas9.
机构信息
Children's Medical Center Research Institute, University of Texas Southwestern Medical Center, Dallas, TX, 75390, USA.
Department of Pediatrics, Harold C. Simmons Comprehensive Cancer Center, and Hamon Center for Regenerative Science and Medicine, University of Texas Southwestern Medical Center, Dallas, TX, 75390, USA.
出版信息
Genome Biol. 2020 Mar 5;21(1):59. doi: 10.1186/s13059-020-01973-w.
The spatiotemporal control of 3D genome is fundamental for gene regulation, yet it remains challenging to profile high-resolution chromatin structure at cis-regulatory elements (CREs). Using C-terminally biotinylated dCas9, endogenous biotin ligases, and pooled sgRNAs, we describe the dCas9-based CAPTURE method for multiplexed analysis of locus-specific chromatin interactions. The redesigned system allows for quantitative analysis of the spatial configuration of a few to hundreds of enhancers or promoters in a single experiment, enabling comparisons across CREs within and between gene clusters. Multiplexed analyses of the spatiotemporal configuration of erythroid super-enhancers and promoter-centric interactions reveal organizational principles of genome structure and function.
三维基因组的时空调控对于基因调控至关重要，但在顺式调控元件（CREs）上进行高分辨率染色质结构分析仍然具有挑战性。我们使用 C 端生物素化的 dCas9、内源性生物素连接酶和汇集的 sgRNA，描述了基于 dCas9 的 CAPTURE 方法，用于对基因座特异性染色质相互作用进行多重分析。重新设计的系统允许在单个实验中对几个到数百个增强子或启动子的空间构象进行定量分析，从而能够在基因簇内和基因簇之间进行 CRE 之间的比较。对红细胞超级增强子和启动子中心相互作用的时空构象的多重分析揭示了基因组结构和功能的组织原则。
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