期刊
MOLECULAR SYSTEMS BIOLOGY
卷 7, 期 -, 页码 -出版社
WILEY
DOI: 10.1038/msb.2010.119
关键词
cell free; in vitro; oscillation; synthetic biology; transcriptional circuits
资金
- ONRYIP [N000140110813]
- NSF [0103002, 0113443, 0608889, 0832824]
- HFSP [RGY0074/2006-C]
- Caltech Center for Biological Circuit Design
- Direct For Computer & Info Scie & Enginr
- Division of Computing and Communication Foundations [0113443] Funding Source: National Science Foundation
- Direct For Computer & Info Scie & Enginr
- Division of Computing and Communication Foundations [832824] Funding Source: National Science Foundation
- Div Of Chem, Bioeng, Env, & Transp Sys
- Directorate For Engineering [0608889, 0103002] Funding Source: National Science Foundation
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. Molecular Systems Biology 7: 465; published online 1 February 2011; doi: 10.1038/msb.2010.119
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