Creating Single-Copy Genetic Circuits

• Main Authors: Choi, Kyeong Rok (Author), Way, Jeffrey C. (Author), Silver, Pamela A. (Author), Lee, Jeongwook (Contributor), Gyoergy, Andras (Contributor), Cameron, David (Contributor), Pyenson, Nora (Contributor), Del Vecchio, Domitilla (Contributor), Collins, James J. (Contributor)
• Other Authors: Institute for Medical Engineering and Science (Contributor), Massachusetts Institute of Technology. Department of Biological Engineering (Contributor), Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science (Contributor), Massachusetts Institute of Technology. Department of Mechanical Engineering (Contributor), Massachusetts Institute of Technology. Synthetic Biology Center (Contributor)
• Format: Article
• Language: English
• Published: Elsevier BV, 2018-08-28T15:55:53Z.
• Subjects: Article
• Online Access: Get fulltext

Synthetic biology is increasingly used to develop sophisticated living devices for basic and applied research. Many of these genetic devices are engineered using multi-copy plasmids, but as the field progresses from proof-of-principle demonstrations to practical applications, it is important to develop single-copy synthetic modules that minimize consumption of cellular resources and can be stably maintained as genomic integrants. Here we use empirical design, mathematical modeling, and iterative construction and testing to build single-copy, bistable toggle switches with improved performance and reduced metabolic load that can be stably integrated into the host genome. Deterministic and stochastic models led us to focus on basal transcription to optimize circuit performance and helped to explain the resulting circuit robustness across a large range of component expression levels. The design parameters developed here provide important guidance for future efforts to convert functional multi-copy gene circuits into optimized single-copy circuits for practical, real-world use.
Defense Threat Reduction Agency (DTRA) (Grant HDTRA1-14-1-006)
United States. Air Force Office of Scientific Research (Grant FA9550-14-1-0060)
United States. Defense Advanced Research Projects Agency (Grant N66001-11-C-4203)
United States. Office of Naval Research (Grant N000141110725)
National Institutes of Health (U.S.) (Grant P50 GM098792)