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Engineering Sensors for Gene Expression Burden.

Alice Boo^1,2, Francesca Ceroni^3,4

¹Department of Bioengineering, Imperial College London, London, UK.

Methods in Molecular Biology (Clifton, N.J.)

|January 6, 2021

Summary

This summary is machine-generated.

We used RNA-sequencing to understand gene expression burden in E. coli, developing sensors and feedback regulators to improve cellular fitness during heterologous gene expression.

Keywords:

Feedback Gene expression burden RNA-seq Sensor Synthetic construct

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Area of Science:

Synthetic Biology
Metabolic Engineering
Molecular Biology

Background:

Gene expression burden negatively impacts cellular growth and predictability in engineered organisms.
Understanding transcriptional responses to burden is crucial for optimizing heterologous gene expression.
Escherichia coli is a key host for biotechnological applications, making burden mitigation important.

Purpose of the Study:

To investigate the transcriptional changes in Escherichia coli under gene expression burden using RNA-sequencing.
To identify early responsive promoters for developing biosensors to detect gene expression burden.
To engineer a feedback regulatory system using a novel sensor to enhance cellular fitness in engineered E. coli.

Main Methods:

RNA-sequencing (RNA-seq) to profile gene expression in E. coli under burden conditions.
Bioinformatic analysis to identify differentially expressed genes and regulatory elements.
Promoter selection and characterization for biosensor development.
Construction and testing of a burden-driven feedback regulator.

Main Results:

RNA-seq identified specific transcriptional profiles associated with gene expression burden in E. coli.
Early responsive promoters were successfully identified and utilized to create burden sensors.
A novel burden-driven feedback regulator was developed, demonstrating improved cellular fitness in engineered E. coli.

Conclusions:

RNA-seq is a powerful tool for dissecting the molecular mechanisms of gene expression burden.
Biosensors derived from early responsive promoters can effectively monitor cellular burden.
Feedback regulation based on burden sensing offers a promising strategy to enhance the performance of engineered microbial systems.