Genomic mining of prokaryotic repressors for orthogonal logic gates
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Voight_Genomic mining.pdf
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Author(s)
Tamsir, Alvin
Clancy, Kevin
Peterson, Todd
Stanton, Brynne C.
Nielsen, Alec Andrew
Voigt, Christopher A.
Date Issued
December 2013
Journal
Nature Chemical Biology
Publisher
Nature Publishing Group
Citation
Stanton, Brynne C, Alec A K Nielsen, Alvin Tamsir, Kevin Clancy, Todd Peterson, and Christopher A Voigt. “Genomic Mining of Prokaryotic Repressors for Orthogonal Logic Gates.” Nat Chem Biol 10, no. 2 (December 8, 2013): 99–105.
Version
Author's final manuscript
Abstract
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[superscript 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.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Synthetic Biology Center
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Creative Commons Attribution-Noncommercial-Share Alike
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DOI of Published Version
https://doi.org/10.1038/nchembio.1411
