Systematic mapping of functional enhancer-promoter connections with CRISPR interference.
Systematic mapping of functional enhancer-promoter connections with CRISPR interference.
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DOI：
10.1126/science.aag2445
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发表时间：
2016-11-11
期刊：
影响因子：
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通讯作者：
Engreitz JM
中科院分区：
文献类型：
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作者：
Fulco CP;Munschauer M;Anyoha R;Munson G;Grossman SR;Perez EM;Kane M;Cleary B;Lander ES;Engreitz JM
哺乳动物中的基因表达受到可影响生理和疾病的非编码元件的调节，但大多数非编码元件的功能和靶基因仍然未知。我们提出了一种使用 CRISPR 干扰 (CRISPRi) 来发现调控元件并识别其靶基因的高通量方法。我们评估了 2 个必需转录因子 MYC 和 GATA1 附近 >1 兆碱基 (Mb) 的序列，并鉴定了 9 个控制基因表达和细胞增殖的远端增强子。染色质状态和染色体构象的定量特征将调节 MYC 的 7 个增强子与其他不调节 MYC 的元件区分开来，这提出了预测增强子-启动子连接性的策略。这种基于 CRISPRi 的方法可用于剖析转录网络并解释非编码遗传变异对人类疾病的贡献。
Gene expression in mammals is regulated by noncoding elements that can impact physiology and disease, yet the functions and target genes of most noncoding elements remain unknown. We present a high-throughput approach that uses CRISPR interference (CRISPRi) to discover regulatory elements and identify their target genes. We assess >1 megabase (Mb) of sequence in the vicinity of 2 essential transcription factors, MYC and GATA1, and identify 9 distal enhancers that control gene expression and cellular proliferation. Quantitative features of chromatin state and chromosome conformation distinguish the 7 enhancers that regulate MYC from other elements that do not, suggesting a strategy for predicting enhancer-promoter connectivity. This CRISPRi-based approach can be applied to dissect transcriptional networks and interpret the contributions of noncoding genetic variation to human disease.