CRISPR Inversion of CTCF Sites Alters Genome Topology and Enhancer/Promoter Function.
CRISPR Inversion of CTCF Sites Alters Genome Topology and Enhancer/Promoter Function.
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CTCF 位点的 CRISPR 倒置改变了基因组拓扑和增强子/启动子功能
DOI：
10.1016/j.cell.2015.07.038
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发表时间：
2015-08-13
期刊：
影响因子：
64.5
通讯作者：
Wu Q
中科院分区：
文献类型：
--
作者：
Guo Y;Xu Q;Canzio D;Shou J;Li J;Gorkin DU;Jung I;Wu H;Zhai Y;Tang Y;Lu Y;Wu Y;Jia Z;Li W;Zhang MQ;Ren B;Krainer AR;Maniatis T;Wu Q
CTCF/cohesin play a central role in insulator function and higher-order chromatin organization of mammalian genomes. Recent studies identified a correlation between the orientation of CTCF-binding sites (CBSs) and chromatin loops. To test the functional significance of this observation, we combined CRISPR/Cas9-based genomic-DNA-fragment editing with chromosome-conformation-capture experiments to show that the location and relative orientations of CBSs determine the specificity of long-range chromatin looping in mammalian genomes, using protocadherin (Pcdh) and β-globin as model genes. Inversion of CBS elements within the Pcdh enhancer reconfigures the topology of chromatin loops between the distal enhancer and target promoters, and alters gene-expression patterns. Thus, although enhancers can function in an orientation-independent manner in reporter assays, in the native chromosome context the orientation of at least some enhancers carrying CBSs can determine both the architecture of topological chromatin domains and enhancer/promoter specificity. The findings reveal how 3D chromosome architecture can be encoded by genome sequence.
