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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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A Multilayer Microfluidic Platform for the Conduction of Prolonged Cell-Free Gene Expression
Ardjan J van der Linden1, Maaruthy Yelleswarapu2, Pascal A Pieters1
1Institute for Complex Molecular Systems, Department of Biomedical Engineering, Computational Biology Group, Eindhoven University of Technology.
Journal of Visualized Experiments : Jove
|October 22, 2019
Summary
Researchers developed a microfluidic device for cell-free synthetic biology, enabling sustained in vitro transcription and translation (IVTT) reactions. This system overcomes limitations of cell-based methods for analyzing complex genetic circuits.
Area of Science:
- Synthetic biology
- Biotechnology
- Microfluidics
Background:
- Cell-based synthetic biology faces limitations in analyzing complex genetic regulatory circuits due to environmental control issues and host interactions.
- In vivo analysis is time-consuming and lacks precise environmental regulation, hindering the study of synthetic gene networks.
- Cell-free systems offer better environmental control for characterizing novel synthetic circuitry.
Purpose of the Study:
- To present a multilayer microfluidic device for sustained in vitro transcription and translation (IVTT) reactions.
- To overcome limitations of batch reactions by enabling resource replenishment and byproduct removal.
- To emulate cellular environments for prolonged investigation of dynamic gene circuit behavior.
Main Methods:
- Fabrication of a multilayer microfluidic device.
- Sustaining in vitro transcription and translation (IVTT) reactions within the microfluidic device.
- Integration of hardware and software for automated IVTT reactions.
Main Results:
- The microfluidic device sustains IVTT reactions for extended durations, emulating cellular environments.
- Resource replenishment and byproduct removal maintain an out-of-equilibrium environment.
- Automated IVTT reactions enable comprehensive analysis of complex network behaviors.
Conclusions:
- The developed microfluidic platform combined with IVTT reactions facilitates the in-depth analysis of complex synthetic genetic regulatory networks.
- This approach advances the understanding of mechanisms regulating cellular processes.
- Enables more robust and controlled characterization of synthetic biology systems.
