Design and analysis of a tunable synchronized oscillator

Area of Science:

• Synthetic Biology
• Systems Biology
• Genetic Engineering

Background:

• Synthetic biology utilizes computational modeling, drawing from nonlinear dynamics and control theory, for designing artificial biological systems.
• Modeling complex genetic circuits presents challenges due to intricate molecular interactions, often leading to oversimplified or intractable models.
• Developing "toy" genetic circuits allows for better experimental control and real-time data acquisition of intracellular states.

Purpose of the Study:

• To design and implement novel genetic circuits capable of synchronized oscillatory gene expression.
• To investigate the enhancement of synchronized transcriptional oscillators using chemically inducible repressors.
• To validate computational models through experimental observation of engineered genetic circuits.

Main Methods:

• Design of two genetic circuits (basic and tunable) for synchronized green fluorescent protein (GFP) expression in Escherichia coli.
• Microscopic verification of the basic circuit's functionality.
• Computational simulations and analysis of high-level visualizations to assess the impact of inducible repressors on oscillator performance.

Main Results:

• Repeated observation of synchronized oscillations in GFP expression within chemically linked bacterial subpopulations.
• Computational predictions indicating that inducible repressors significantly expand the operational parameter space for oscillations.
• Demonstrated tunability of oscillation frequency through the introduction of independently inducible repressors.

Conclusions:

• The developed genetic circuits serve as valuable research tools for studying synchronized transcriptional feedback loops.
• Abstract visualizations aid in uncovering non-linear dynamics within complex computational models.
• The findings highlight the potential for enhanced control and broader applicability of synthetic genetic oscillators.