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Updated: Jun 10, 2026

Generation of Heterogeneous Drug Gradients Across Cancer Populations on a Microfluidic Evolution Accelerator for Real-Time Observation

Generation of Heterogeneous Drug Gradients Across Cancer Populations on a Microfluidic Evolution Accelerator for Real-Time Observation

Published on: September 19, 2019

Microfluidic compartmentalized directed evolution.

Brian M Paegel¹, Gerald F Joyce

¹Department of Chemistry, The Scripps Research Institute, 130 Scripps Way, Jupiter, FL 33458, USA. briandna@scripps.edu

Chemistry & Biology

|July 28, 2010

Summary

This summary is machine-generated.

A new microfluidic device creates uniform droplets for directed evolution, enabling the selection of RNA enzymes resistant to neomycin. This high-throughput method enhances biochemical reagent stability and evolutionary efficiency.

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Area of Science:

Biochemistry
Molecular Biology
Biotechnology

Background:

Conventional bulk emulsification for directed evolution yields nonuniform droplets, potentially damaging reagents.
Microfluidic technologies offer precise control over droplet generation for biological applications.

Purpose of the Study:

To develop a microfluidic emulsification circuit for generating uniform water-in-oil droplets.
To utilize this device for directed evolution of RNA enzymes with enhanced properties.

Main Methods:

A microfluidic circuit with a radial array of aqueous nozzles intersecting an oil flow channel was designed.
The device achieved high throughput (10^7-10^8 droplets/hour) and uniform droplet size (21.9 ± 0.8 µm radius).
RNA enzymes with RNA ligase activity were evolved for neomycin resistance through multiple rounds of selective amplification.

Main Results:

The microfluidic device successfully produced highly uniform droplets at high throughput.
Directed evolution yielded RNA enzymes that gained resistance to neomycin inhibition.
Some evolved enzymes exhibited neomycin-dependent optimal activity, indicating significant functional changes.

Conclusions:

Microfluidic emulsification provides a superior method for generating compartments in directed evolution compared to bulk techniques.
This technology facilitates efficient evolution of biomolecules with desired functional traits.
The evolved RNA enzymes demonstrate potential for novel applications in molecular biology and biotechnology.