Orthogonal intercellular signaling for programmed spatial behavior.
Orthogonal intercellular signaling for programmed spatial behavior.
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DOI：
10.15252/msb.20156590
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发表时间：
2016-01-25
影响因子：
9.9
通讯作者：
Haseloff J
中科院分区：
文献类型：
--
作者：
Grant PK;Dalchau N;Brown JR;Federici F;Rudge TJ;Yordanov B;Patange O;Phillips A;Haseloff J
双向细胞间信号传导是多细胞生物的一个基本特征，复杂生物系统的工程构建将需要具有最小串扰的多种细胞间信号传导途径。天然的群体感应系统为细胞通讯提供了组件，但其使用往往受到信号串扰的限制。我们利用酰基高丝氨酸内酯信号系统建立了新的用于细胞 - 细胞通讯的正交系统。在不同受体蛋白表达情况下的定量测量使我们能够区分3 - 氧代 - C6 - 和3 - 氧代 - C12 - 高丝氨酸内酯、同源受体蛋白以及DNA启动子之间不同类型的串扰。对启动子序列进行突变可将与异源受体蛋白的相互作用降至最低。我们利用实验数据对信号串扰的计算模型进行参数化，并估计受体蛋白水平对信号串扰的影响。我们利用该模型预测受体蛋白的最佳表达水平，以创建一种有效的双通道细胞通讯装置。一种新型空间测定方法的建立使得能够通过这些可扩散信号测量几何受限的细胞群体之间的相互作用。我们构建了能够通过离散细胞群体的信号诱导、通讯和响应循环介导长距离信号传播的中继装置。这项工作证明了能够系统地减少细胞间信号传导系统内的串扰，并利用这些系统对细胞群体进行复杂的时空模式构建。
Bidirectional intercellular signaling is an essential feature of multicellular organisms, and the engineering of complex biological systems will require multiple pathways for intercellular signaling with minimal crosstalk. Natural quorum‐sensing systems provide components for cell communication, but their use is often constrained by signal crosstalk. We have established new orthogonal systems for cell–cell communication using acyl homoserine lactone signaling systems. Quantitative measurements in contexts of differing receiver protein expression allowed us to separate different types of crosstalk between 3‐oxo‐C6‐ and 3‐oxo‐C12‐homoserine lactones, cognate receiver proteins, and DNA promoters. Mutating promoter sequences minimized interactions with heterologous receiver proteins. We used experimental data to parameterize a computational model for signal crosstalk and to estimate the effect of receiver protein levels on signal crosstalk. We used this model to predict optimal expression levels for receiver proteins, to create an effective two‐channel cell communication device. Establishment of a novel spatial assay allowed measurement of interactions between geometrically constrained cell populations via these diffusible signals. We built relay devices capable of long‐range signal propagation mediated by cycles of signal induction, communication and response by discrete cell populations. This work demonstrates the ability to systematically reduce crosstalk within intercellular signaling systems and to use these systems to engineer complex spatiotemporal patterning in cell populations.