Population-level amplification of gene regulation by programmable gene transfer

Area of Science:

• Synthetic Biology
• Microbial Engineering
• Genetic Circuit Design

Background:

• Single-cell gene regulation often limits dynamic range and control.
• Population-level mechanisms, like immune responses, offer robust control strategies.
• Existing synthetic biology tools struggle with scalable, dynamic population-wide gene regulation.

Purpose of the Study:

• To develop a programmable system for dynamic, population-level gene regulation in bacteria.
• To leverage plasmid-mediated gene transfer for amplifying single-cell regulatory signals.
• To expand the dynamic range and orthogonality of gene expression in engineered microbial communities.

Main Methods:

• Utilized Cas9 endonuclease, F conjugation machinery, and antibiotic selection to control plasmid dynamics.
• Regulated plasmid loss rate, transfer rate, and fitness effects to modulate plasmid-carrying cell fraction.
• Implemented a plasmid-mediated gene transfer system for programmable population control.

Main Results:

• Demonstrated dynamic control and amplification of gene regulation in bacterial populations.
• Successfully modulated the fraction of plasmid-carrying cells, acting as an amplification factor.
• Achieved expanded dynamic range and orthogonal control of gene expression across cell populations.

Conclusions:

• The developed platform enables dynamic regulation of gene expression in engineered microbial communities.
• Plasmid-mediated gene transfer provides a versatile strategy for population-level synthetic biology applications.
• This approach enhances the scalability and robustness of engineered biological systems.