Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric Receptors.
Engineering Synthetic Signaling Pathways with Programmable dCas9-Based Chimeric Receptors.
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DOI：
10.1016/j.celrep.2017.08.044
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发表时间：
2017-09-12
期刊：
影响因子：
8.8
通讯作者：
Fulga TA
中科院分区：
文献类型：
--
作者：
Baeumler TA;Ahmed AA;Fulga TA
合成受体为产生能够监测环境、感知特定输入信号和执行不同定制反应程序的设计细胞提供了强大的实验工具。为了推进细胞工程的前景，我们开发了一类嵌合受体，它集成了一个高度可编程和便携式的核酸酶缺陷CRISPR/Cas9(DCas9)信号转导模块。我们证明了核心的dCas9合成受体(dCas9-synR)结构可以很容易地适应各种类型的天然胞外结构域支架，将它们的自然输入与正交输出函数联系起来。重要的是，这些受体实现了严格的OFF/ON状态转换特征，表现出激动剂介导的剂量依赖的激活，并且可以编程将特定的疾病标记物与不同的、治疗相关的多基因表达电路偶联。这里开发的模块化dCas9-synR平台为设计下一代合成受体提供了通用的蓝图，这将使细胞工程中实现高度复杂的组合功能成为可能。基于高度可编程的Split-dCas9信号转导模块的dCas9-SynR核心架构可以跨多种感应域进行标准化集成基本dCas9-SynR架构中的用户控制与门dCas9-SynR使复杂的组合治疗反应能够使用高度可编程的基于Split-dCas9的信号转导模块来实施，Baeumler等人。已经创造了一类新型的合成受体(dCas9-synr)，能够将生物相关的输入信号与定制的用户定义的输出响应程序的直接激活相耦合。DCas9-synRs扩大了细胞工程在研究和治疗应用方面的前景。
Synthetic receptors provide a powerful experimental tool for generation of designer cells capable of monitoring the environment, sensing specific input signals, and executing diverse custom response programs. To advance the promise of cellular engineering, we have developed a class of chimeric receptors that integrate a highly programmable and portable nuclease-deficient CRISPR/Cas9 (dCas9) signal transduction module. We demonstrate that the core dCas9 synthetic receptor (dCas9-synR) architecture can be readily adapted to various classes of native ectodomain scaffolds, linking their natural inputs with orthogonal output functions. Importantly, these receptors achieved stringent OFF/ON state transition characteristics, showed agonist-mediated dose-dependent activation, and could be programmed to couple specific disease markers with diverse, therapeutically relevant multi-gene expression circuits. The modular dCas9-synR platform developed here provides a generalizable blueprint for designing next generations of synthetic receptors, which will enable the implementation of highly complex combinatorial functions in cellular engineering. dCas9-synRs based on a highly programmable split-dCas9 signal transduction module Core architecture can be standardized across multiple classes of sensing domains Integration of user-controlled AND gates in the basic dCas9-synR architecture dCas9-synRs enable implementation of complex combinatorial therapeutic responses Using a highly programmable split-dCas9-based signal transduction module, Baeumler et al. have created a novel class of synthetic receptors (dCas9-synRs) capable of coupling biologically relevant input signals with the direct activation of custom user-defined output response programs. dCas9-synRs expand the promise of cellular engineering for research and therapeutic applications.
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DOI：
10.1038/nrm2904
发表时间：
2010-06
期刊：
Nature reviews. Molecular cell biology
影响因子：
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作者：
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影响因子：
64.8
作者：
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发表时间：
2014-02
期刊：
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影响因子：
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影响因子：
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作者：
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