Journal
NATURE CHEMICAL BIOLOGY
Volume 10, Issue 2, Pages 99-105Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nchembio.1411
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Funding
- Department of Defense, Air Force Office of Scientific Research, National Defense Science and Engineering Graduate Fellowship, 32 CFR 168a [FA9550-11-C-0028]
- Life Technologies, Defense Advanced Research Projects Agency Chronicle of Lineage Indicative of Origins (DARPA CLIO) [N66001-12-C-4016]
- Office of Naval Research [N00014-13-1-0074]
- US National Institutes of Health [GM095765]
- US National Science Foundation Synthetic Biology Engineering Research Center (SynBERC) [SA5284-11210]
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Genetic circuits perform computational operations based on interactions between freely diffusing molecules within a cell. When transcription factors are combined to build a circuit, unintended interactions can disrupt its function. Here, we apply 'part mining' to build a library of 73 TetR-family repressors gleaned from prokaryotic genomes. The operators of a subset were determined using an in vitro method, and this information was used to build synthetic promoters. The promoters and repressors were screened for cross-reactions. Of these, 16 were identified that both strongly repress their cognate promoter (5- to 207-fold) and exhibit minimal interactions with other promoters. Each repressor-promoter pair was converted to a NOT gate and characterized. Used as a set of 16 NOT/NOR gates, there are >10(54) circuits that could be built by changing the pattern of input and output promoters. This represents a large set of compatible gates that can be used to construct user-defined circuits.
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