Creating Single-Copy Genetic Circuits
Name
nihms855180.pdf
Size
1.46 MB
Format
Adobe PDF
Checksum (MD5)
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Author(s)
Choi, Kyeong Rok
Way, Jeffrey C.
Silver, Pamela A.
Lee, Jeongwook
Gyoergy, Andras
Cameron, David
Pyenson, Nora
Del Vecchio, Domitilla
Collins, James J.
Date Issued
July 2016
Journal
Molecular Cell
Publisher
Elsevier BV
Citation
Lee, Jeong Wook et al. “Creating Single-Copy Genetic Circuits.” Molecular Cell 63, 2 (July 2016): 329–336 © 2016 Elsevier Inc
Version
Final published version
Abstract
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.
MIT Department
Institute for Medical Engineering and Science
Massachusetts Institute of Technology. Department of Biological Engineering
Massachusetts Institute of Technology. Department of Electrical Engineering and Computer Science
Massachusetts Institute of Technology. Department of Mechanical Engineering
Massachusetts Institute of Technology. Synthetic Biology Center
Terms of Use
Creative Commons Attribution-NonCommercial-NoDerivs License
Persistent DSpace Link
DOI of Published Version
https://doi.org/10.1016/J.MOLCEL.2016.06.006
