Dynamic control of endogenous metabolism with combinatorial logic circuits

Area of Science:

• Synthetic Biology
• Metabolic Engineering
• Biotechnology

Background:

• Controlling gene expression is crucial for real-time metabolic control and cellular response staging in bioprocesses.
• Small molecule inducers are impractical for large-scale bioprocesses, and dynamic gene circuits are challenging to design.
• Dynamic gene regulation is needed for optimizing bioprocesses and achieving desired cellular behaviors.

Purpose of the Study:

• To demonstrate the integration of sensors into combinatorial logic circuits for dynamic gene regulation in Escherichia coli.
• To implement temporal control over gene expression during a bioprocess using engineered logic circuits.
• To enable customizable dynamic gene regulation for metabolic control and by-product management.

Main Methods:

• Engineered three Escherichia coli sensors responding to glucose, dissolved oxygen, and acetate accumulation.
• Integrated sensors into combinatorial logic circuits to implement temporal control over an 18-hour period.
• Regulated endogenous enzymes at transcriptional (CRISPRi) and post-translational (targeted proteolysis) levels.

Main Results:

• Successfully implemented logic circuits for temporal control of gene expression in Escherichia coli.
• Demonstrated dynamic regulation of acetate production by controlling endogenous genes (pta or poxB).
• Showcased customizable dynamic gene regulation using simple circuit designs.

Conclusions:

• Engineered logic circuits with integrated sensors enable precise temporal control of gene expression in bioprocesses.
• This approach offers a flexible platform for dynamic metabolic control and optimization of cellular functions.
• The developed circuits provide a foundation for advanced synthetic biology applications in biotechnology.