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Published on: January 7, 2019
Development of Cell-Based Sentinels for Nitric Oxide: Ensuring Marker Expression and Unimodality
Ryan McKay1,2, Pricila Hauk1,2, David Quan1,2
1Fischell Department of Bioengineering , University of Maryland , College Park , Maryland 20742 , United States.
Researchers engineered bacteria to detect nitric oxide (NO), a biomarker for Crohn's disease. These novel biosensors offer a potential non-invasive diagnostic tool, contrasting with traditional methods like colonoscopies.
Area of Science:
- Synthetic Biology
- Microbial Engineering
- Biomarker Detection
Background:
- Nitric oxide (NO) is an elevated biomarker in intestinal inflammation, characteristic of Crohn's disease (CD).
- Current diagnostic methods for CD, such as colonoscopies and biopsies, are invasive and burdensome.
Purpose of the Study:
- To develop a novel bacterial biosensor capable of detecting nitric oxide (NO).
- To engineer a responsive genetic circuit for a potential ingestible diagnostic tool for Crohn's disease.
Main Methods:
- Construction of a dual-plasmid system with an NO-responsive promoter and T7 Polymerase (T7Pol) amplification.
- Optimization of T7Pol activity and translational control to balance cellular transcriptional load.
- Transformation of commensal E. coli Nissle with the genetic circuitry for in vivo application.
Main Results:
- Generated "sentinel" bacteria exhibiting a homogeneous and strong fluorescent response to nitric oxide.
- Successfully engineered a genetic circuit that balances transcriptional load for optimal biosensor performance.
- Demonstrated proof-of-concept for an ingestible, cell-based sensor for detecting elevated intestinal NO in Crohn's disease.
Conclusions:
- The developed NO-responsive bacterial biosensors offer a simple and potentially non-invasive diagnostic approach for Crohn's disease.
- This microbial engineering strategy provides a foundation for ingestible cell-based diagnostic tools.
- The system highlights the potential of synthetic biology for creating advanced diagnostic solutions.
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