title: Engineering orthogonal dual transcription factors for multi-input synthetic promoters.
abstract: Synthetic biology has seen an explosive growth in the capability of engineering artificial gene circuits from transcription factors (TFs), particularly in bacteria. However, most artificial networks still employ the same core set of TFs (for example LacI, TetR and cI). The TFs mostly function via repression and it is difficult to integrate multiple inputs in promoter logic. Here we present to our knowledge the first set of dual activator-repressor switches for orthogonal logic gates, based on bacteriophage λ cI variants and multi-input promoter architectures. Our toolkit contains 12 TFs, flexibly operating as activators, repressors, dual activator-repressors or dual repressor-repressors, on up to 270 synthetic promoters. To engineer non cross-reacting cI variants, we design a new M13 phagemid-based system for the directed evolution of biomolecules. Because cI is used in so many synthetic biology projects, the new set of variants will easily slot into the existing projects of other groups, greatly expanding current engineering capacities.
authors.affiliation: Department of Life Sciences, Imperial College London, London SW7 2AZ, UK.
authors.forename: Andreas K
authors.initials: AK
authors.lastname: Brödel
authors.affiliation: School of Life Sciences, University of Warwick, Coventry CV4 7AL, UK.|Institute of Systems and Synthetic Biology, Genopole, CNRS, Université d'Évry, 91030 Évry, France.
authors.forename: Alfonso
authors.identifier: 0000-0002-6313-9689
authors.initials: A
authors.lastname: Jaramillo
authors.affiliation: Department of Life Sciences, Imperial College London, London SW7 2AZ, UK.
authors.forename: Mark
authors.identifier: 0000-0002-4652-0365
authors.initials: M
authors.lastname: Isalan
mesh_terms: D001435:Bacteriophages; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA; D006801:Humans; D010957:Plasmids; D011401:Promoter Regions, Genetic; D015202:Protein Engineering; D014157:Transcription Factors / Q000737:chemistry / Q000235:genetics*
chemical_list: D014157:Transcription Factors; D004247:DNA