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On a Robust, Sensitive Cell-Free Method for Pseudomonas Sensing and Quantification in Microfluidic Templated
Jong Seto1,2
1Department of Bioengineering and Therapeutic Sciences, University of California at San Francisco and California, Institute for Quantitative Biosciences (QB3), 1700 4th Street, Byers Hall #303, San Francisco, CA 94158, USA. JMSeto@lbl.gov.
Micromachines
|August 3, 2019
Summary
This study presents a cell-free platform using hydrogel beads for biological functions without cells. The system enables robust sensing of quorum-sensing molecules, crucial for pathogen detection.
Area of Science:
- Synthetic Biology
- Biotechnology
- Microfluidics
Background:
- Cell-free systems offer a way to perform biological functions like transcription and translation outside of living cells.
- This approach bypasses the need for optimal cell growth conditions and complex cellular machinery.
- It allows for the precise engineering of synthetic biological circuits.
Purpose of the Study:
- To develop a cell-free platform for biologically relevant functions.
- To demonstrate a robust and portable sensing application for quorum-sensing molecules.
- To create an adaptable hydrogel bead system for remote environmental sensing.
Main Methods:
- Utilizing droplet microfluidics to encapsulate cell-free lysates within hydrogel beads.
- Reconstituting in vitro transcription and translation by isolating key biological components.
- Developing an agarose bead platform for sensing specific molecules.
Main Results:
- Demonstrated a cell-free platform capable of performing biological functions without cells.
- Successfully applied the platform for sensing the quorum-sensing molecule N-3-oxododecanoyl homoserine lactone (3OC12HSL).
- Developed a robust, portable, and substrate-independent sensing system.
Conclusions:
- Cell-free systems integrated into hydrogel beads provide a versatile platform for synthetic biology.
- This technology enables adaptable and programmable sensing for various biological and chemical analytes.
- The platform is suitable for delivery and activation in remote or challenging environments.
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