Synthetic in vitro transcriptional oscillators.
Synthetic in vitro transcriptional oscillators.
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DOI：
10.1038/msb.2010.119
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发表时间：
2011-02-01
影响因子：
9.9
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中科院分区：
文献类型：
--
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简单组件中的合成生化回路的构建阐明了化学中如何出现复杂行为，并为将来的生物学技术奠定了基础。遗传调节网络的简化类似物，体外转录电路，为系统构建任意电路提供了一个模块化平台，仅需要两种必需酶，噬菌体T7 RNA聚合酶和埃斯切里虫大肠杆菌核糖核酸酶H，以产生和脱粒RNA信号。在这项研究中，我们在体外设计和实验证明了三个转录振荡器。首先，由两个开关组成的负反馈振荡器，由兴奋性和抑制性RNA信号调节，最多显示了五个完整的周期。为了证明模块化并进一步探索设计空间，添加了一个正反馈回路，该回路调节并扩展了振荡状态。最后，构建和分析了三个开关环振荡器。数学建模指导了设计过程，确定了可能产生振荡的实验条件，并解释了该系统对短降解产物干扰的强大反应。合成转录振荡器可能证明对于生化电路设计原理的系统探索以及控制纳米级设备和在人造细胞内的过程中的过程很有价值。
The construction of synthetic biochemical circuits from simple components illuminates how complex behaviors can arise in chemistry and builds a foundation for future biological technologies. A simplified analog of genetic regulatory networks, in vitro transcriptional circuits, provides a modular platform for the systematic construction of arbitrary circuits and requires only two essential enzymes, bacteriophage T7 RNA polymerase and Escherichia coli ribonuclease H, to produce and degrade RNA signals. In this study, we design and experimentally demonstrate three transcriptional oscillators in vitro. First, a negative feedback oscillator comprising two switches, regulated by excitatory and inhibitory RNA signals, showed up to five complete cycles. To demonstrate modularity and to explore the design space further, a positive-feedback loop was added that modulates and extends the oscillatory regime. Finally, a three-switch ring oscillator was constructed and analyzed. Mathematical modeling guided the design process, identified experimental conditions likely to yield oscillations, and explained the system's robust response to interference by short degradation products. Synthetic transcriptional oscillators could prove valuable for systematic exploration of biochemical circuit design principles and for controlling nanoscale devices and orchestrating processes within artificial cells.
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