Genomic mining of prokaryotic repressors for orthogonal logic gates
Author(s)
Tamsir, Alvin; Clancy, Kevin; Peterson, Todd; Stanton, Brynne C.; Nielsen, Alec Andrew; Voigt, Christopher A.; ... Show more Show less![Thumbnail](/bitstream/handle/1721.1/99526/Voight_Genomic%20mining.pdf.jpg?sequence=4&isAllowed=y)
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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[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.
Date issued
2013-12Department
Massachusetts Institute of Technology. Department of Biological Engineering; Massachusetts Institute of Technology. Synthetic Biology CenterJournal
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
ISSN
1552-4450
1552-4469