Journal
NATURE BIOTECHNOLOGY
Volume 27, Issue 10, Pages 946-U112Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nbt.1568
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Funding
- Pew and Packard Foundations
- Office of Naval Research
- National Institutes of Health (NIH) [EY016546, AI067699]
- NSF [BES-0547637]
- TeraGrid [TG-MCB080126T]
- NSF Graduate Research Fellowship
- American Society for Engineering Education National Defense Science and Engineering Graduate Fellowship
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Microbial engineering often requires fine control over protein expression-for example, to connect genetic circuits(1-7) or control flux through a metabolic pathway(8-13). To circumvent the need for trial and error optimization, we developed a predictive method for designing synthetic ribosome binding sites, enabling a rational control over the protein expression level. Experimental validation of >100 predictions in Escherichia coli showed that the method is accurate to within a factor of 2.3 over a range of 100,000-fold. The design method also correctly predicted that reusing identical ribosome binding site sequences in different genetic contexts can result in different protein expression levels. We demonstrate the method's utility by rationally optimizing protein expression to connect a genetic sensor to a synthetic circuit. The proposed forward engineering approach should accelerate the construction and systematic optimization of large genetic systems.
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