CRISPR-Based Editing Reveals Edge-Specific Effects in Biological Networks
CRISPR-Based Editing Reveals Edge-Specific Effects in Biological Networks
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DOI：
10.1089/crispr.2018.0018
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发表时间：
2018-08-01
期刊：
影响因子：
3.7
通讯作者：
Bleris, Leonidas
中科院分区：
文献类型：
--
作者：
Li, Yi;Nowak, Chance M.;Bleris, Leonidas
揭示生物网络的特性是理解正常和疾病细胞表型的核心。网络由功能元素（节点）组成，这些功能元素（节点）形成各种不同的连接（边），每个节点是多个边的枢纽。在此，与以节点为中心的网络扰动和分析方法相比，我们提出了一种基于crispr的高通量方法来描述网络边缘的作用。使用针对71个基因中的93个miRNA靶点的文库消蚀网络边缘，揭示了在压力下控制细胞生长和存活的众多边缘，其重要性各不相同。为了比较去除生物网络中节点和边缘的影响，我们剖析了一个特定的p53-microRNA通路。我们发现，从抗凋亡基因BCL2中去除miR-34a的靶位点会以p53依赖的方式使细胞对miR-34a的异位递送脱敏。综上所述，我们证明了网络边缘对生物网络的功能和稳定性至关重要。我们的研究结果通过边缘消融引入了一种新的基因筛选机会，并突出了生物网络分析的新维度。
Unraveling the properties of biological networks is central to understanding both normal and disease cellular phenotypes. Networks consist of functional elements (nodes) that form a variety of diverse connections (edges), with each node being a hub for multiple edges. Herein, in contrast to node-centric network perturbation and analysis approaches, we present a high-throughput CRISPR-based methodology for delineating the role of network edges. Ablation of network edges using a library targeting 93 miRNA target sites in 71 genes reveals numerous edges that control, with variable importance, cellular growth and survival under stress. To compare the impact of removing nodes versus edges in a biological network, we dissect a specific p53-microRNA pathway. We show that removal of the miR-34a target site from the anti-apoptotic gene BCL2 desensitizes the cell to ectopic delivery of miR-34a in a p53-dependent manner. In summary, we demonstrate that network edges are critical to the function and stability of biological networks. Our results introduce a novel genetic screening opportunity via edge ablation and highlight a new dimension in biological network analysis.