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Rapid cell-free forward engineering of novel genetic ring oscillators
Abstract While complex dynamic biological networks …
While complex dynamic biological networks control gene expression in all living organisms, the forward engineering of comparable synthetic networks remains challenging. The current paradigm of characterizing synthetic networks in cells results in lengthy design-build-test cycles, minimal data collection, and poor quantitative characterization. Cell-free systems are appealing alternative environments, but it remains questionable whether biological networks behave similarly in cell-free systems and in cells. We characterized in a cell-free system the 'repressilator,' a three-node synthetic oscillator. We then engineered novel three, four, and five-gene ring architectures, from characterization of circuit components to rapid analysis of complete networks. When implemented in cells, our novel 3-node networks produced population-wide synchronized oscillations and 95% of 5-node oscillator cells oscillated for up to 72 hours. Oscillation periods in cells matched the cell-free system results for all networks tested. An alternate forward engineering paradigm using cell-free systems can thus accurately capture cellular behavior.
thus accurately capture cellular behavior.  +
Authors Henrike Niederholtmeyer, Zachary Sun, Yutaka Hori, Enoch Yeung, Amanda Verpoorte, Richard M Murray and Sebastian J Maerkl  +
Flags Biocircuits  +
Funding Biomolecular Breadboards for Prototyping and Debugging Synthetic Biocircuits +
ID 2015i  +
Source ''eLife'' 2015;10.7554/eLife.09771  +
Tag nie15-elife  +
Title Rapid cell-free forward engineering of novel genetic ring oscillators +
Type Journal paper  +
Categories Papers
Modification date
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26 January 2019 15:55:15  +
URL
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http://elifesciences.org/content/early/2015/10/02/eLife.09771  +
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Rapid cell-free forward engineering of novel genetic ring oscillators + Title
 

 

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