On a Robust, Sensitive Cell-Free Method for Pseudomonas Sensing and Quantification in Microfluidic Templated Hydrogels

Micromachines
Jong Seto

Abstract

Through the use of droplet microfluidics to integrate cell-free activity into inert hydrogel beads, we have developed a platform that can perform biologically relevant functions without the need for cells. Specifically, cell-free lysates serve a utility in performing cellular functions and providing biologically relevant metabolic products without requiring the optimal biological conditions for cell growth and proliferation. By teasing out specific biological components that enable transcription and translation to occur, these cell-like functions can be reconstituted in vitro without requiring the entire cell and milieu of cellular organelles. This enables the optimization of synthetic biological circuits, either by concentration or logic switches, simply through the addition or removal of genetic components (plasmids, inducers, or repressors) of regulatory elements. Here, we demonstrate an application of cell-free processes that is robust and portable, independent of a substrate, to apply for sensing and reporting functions of a quorum-sensing molecule N-3-oxododecanoyl homoserine lactone (3OC12HSL) found crucial for pathological Pseudomonas aeruginosa infection. We develop an agarose bead platform that is easily adaptable and...Continue Reading

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Citations

Jul 16, 2020·Biotechnology Journal·Liyuan ZhangYuan Lu
Dec 31, 2020·Micromachines·Ai Qun LiuYi Zhang
Aug 7, 2021·Frontiers in Bioengineering and Biotechnology·Megan A McSweeney, Mark P Styczynski

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Methods Mentioned

BETA
biosensor
Chip
FACS
fluorescence microscopy
fluorescence-activated cell sorting
fluorescence imaging

Software Mentioned

Avizo 3D
SoftWorX
FlowJo
ImageJ

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