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A Multilayer Microfluidic Platform for the Conduction of Prolonged Cell-Free Gene Expression
Published on: October 6, 2019
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Enabling complex genetic circuits to respond to extrinsic environmental signals
Allison Hoynes-O'Connor1, Tatenda Shopera1, Kristina Hinman1
1Department of Energy, Environmental and Chemical Engineering, Washington University in St. Louis, St. Louis 63130, One Brookings Dr., Box 1180, Missouri.
Biotechnology and Bioengineering
|March 7, 2017
Summary
Researchers engineered novel genetic circuits that respond to temperature and pH changes. These synthetic biological tools, featuring AND and NAND logic gates, are crucial for advancing metabolic engineering and biotechnological applications.
Area of Science:
- Synthetic biology
- Genetic circuit engineering
- Biotechnology
Background:
- Genetic circuits are essential for metabolic engineering, medical, and environmental applications.
- Implementing sensors for extrinsic and intrinsic signals is critical for functional genetic circuits.
- Existing genetic circuits lack responsiveness to key environmental factors like temperature and pH.
Purpose of the Study:
- To construct the first AND and NAND logic gates responsive to temperature and pH.
- To characterize and engineer pH- and temperature-responsive promoters.
- To integrate these components into functional genetic circuits for real-world applications.
Main Methods:
- Extraction and characterization of pH- and temperature-responsive promoters from E. coli.
- Modification of promoters for genetic circuit applications.
- Integration of promoters with Salmonella typhimurium type III secretion system components.
- Construction of AND gates and subsequent inversion using antisense RNA to create NAND gates.
Main Results:
- Successfully constructed AND gates with up to 23-fold change in response to temperature and pH.
- Developed NAND gates with a significant 1168-fold change by inverting AND gate logic.
- Demonstrated the first complex pH- and temperature-responsive genetic circuits.
Conclusions:
- The developed genetic circuits represent a significant advancement in synthetic biology.
- These circuits provide a foundation for future applications requiring environmental responsiveness.
- This work paves the way for sophisticated genetic tools in biotechnology and medicine.
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