Engineering orthogonal dual transcription factors for multi-input synthetic promoters
File(s)ncomms13858.pdf (648.67 KB)
Published version
OA Location
Author(s)
Broedel, AK
Jaramillo, A
Isalan, M
Type
Journal Article
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.
Date Issued
2016-12-16
Date Acceptance
2016-11-04
Citation
Nature Communications, 2016, 7, pp.1-9
ISSN
2041-1723
Publisher
Nature Publishing Group
Start Page
1
End Page
9
Journal / Book Title
Nature Communications
Volume
7
Copyright Statement
© 2016 The Author(s). This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
Sponsor
Wellcome Trust
Wellcome Trust
Commission of the European Communities
Identifier
https://www.nature.com/articles/ncomms13858
Grant Number
102944/Z/13/Z
102944/Z/13/Z
610730
Subjects
Science & Technology
Multidisciplinary Sciences
Science & Technology - Other Topics
LAMBDA-REPRESSOR
CONTINUOUS EVOLUTION
DIRECTED EVOLUTION
GENETIC CIRCUIT
PROTEIN
DESIGN
SYSTEM
ACTIVATION
PRINCIPLES
RESISTANCE
Bacteriophages
Base Sequence
Cell Line
Cloning, Molecular
DNA
Humans
Plasmids
Promoter Regions, Genetic
Protein Engineering
Transcription Factors
Cell Line
Humans
Bacteriophages
Transcription Factors
DNA
Cloning, Molecular
Protein Engineering
Base Sequence
Plasmids
Promoter Regions, Genetic
Publication Status
Published
Article Number
13858
Date Publish Online
2016-12-16