Orthogonal Genetic Regulation in Human Cells Using Chemically Induced CRISPR/Cas9 Activators
Orthogonal Genetic Regulation in Human Cells Using Chemically Induced CRISPR/Cas9 Activators
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DOI：
10.1021/acssynbio.6b00313
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发表时间：
2017-04-01
影响因子：
4.7
通讯作者：
Zhao, Huimin
中科院分区：
文献类型：
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作者：
Bao, Zehua;Jain, Surbhi;Zhao, Huimin
多个基因以时间依赖的方式协同作用控制复杂的细胞表型，然而基因表达的时间调控仅限于单基因水平，这限制了我们控制高阶基因网络和理解多重遗传扰动后果的能力。在这里，我们开发了一个系统的多个基因的时间调节。该系统结合了CRISPR/Cas9激活剂用于多个基因的正交靶向的简单性和化学诱导二聚化（CID）蛋白用于CRISPR/Cas9激活剂功能的时间控制的正交性。在人类细胞中，这些转录激活因子以配体依赖性方式在最小背景下同时激活多个基因并正交调节不同基因。我们设想，我们的系统将使扰动高阶基因网络具有高的时间分辨率，并加快我们的理解基因在一个复杂的生物环境中的相互作用。
The concerted action of multiple genes in a time-dependent manner controls complex cellular phenotypes, yet the temporal regulation of gene expressions is restricted on a single-gene level, which limits our ability to control higher-order gene networks and understand the consequences of multiplex genetic perturbations. Here we developed a system for temporal regulation of multiple genes. This system combines the simplicity of CRISPR/Cas9 activators for orthogonal targeting of multiple genes and the orthogonality of chemically induced dimerizing (CID) proteins for temporal control of CRISPR/Cas9 activator function. In human cells, these transcription activators exerted simultaneous activation of multiple genes and orthogonal regulation of different genes in a ligand-dependent manner with minimal background. We envision that our system will enable the perturbation of higher-order gene networks with high temporal resolution and accelerate our understanding of gene gene interactions in a complex biological setting.