期刊
SCIENCE
卷 334, 期 6063, 页码 1716-1719出版社
AMER ASSOC ADVANCEMENT SCIENCE
DOI: 10.1126/science.1212209
关键词
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资金
- Synthetic Biology Engineering Research Center (SynBERC) through NSF [0540879]
- JBEI through Lawrence Berkeley National Laboratory [DE-AC02-05CH11231]
- JBEI through U.S. Department of Energy [DE-AC02-05CH11231]
- Jane Coffin Childs Postdoctoral Fellowship
The models and simulation tools available to design functionally complex synthetic biological devices are very limited. We formulated a design-driven approach that used mechanistic modeling and kinetic RNA folding simulations to engineer RNA-regulated genetic devices that control gene expression. Ribozyme and metabolite-controlled, aptazyme-regulated expression devices with quantitatively predictable functions were assembled from components characterized in vitro, in vivo, and in silico. The models and design strategy were verified by constructing 28 Escherichia coli expression devices that gave excellent quantitative agreement between the predicted and measured gene expression levels (r = 0.94). These technologies were applied to engineer RNA-regulated controls in metabolic pathways. More broadly, we provide a framework for studying RNA functions and illustrate the potential for the use of biochemical and biophysical modeling to develop biological design methods.
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