Modular and tunable biological feedback control using a de novo protein switch

Area of Science:

• Synthetic Biology
• Protein Engineering
• Molecular Biology

Background:

• De novo-designed proteins offer versatile tools for synthetic biology, complementing engineered natural proteins.
• The degronLOCKR system, based on LOCKR technology, acts as a switch to degrade target proteins in vivo upon small peptide induction.

Purpose of the Study:

• To implement feedback control in endogenous signaling pathways and synthetic gene circuits using the degronLOCKR system.
• To demonstrate the plug-and-play capability of degronLOCKR for rewiring cellular pathways and engineering synthetic circuits.
• To evaluate the feedback capabilities and operational range of degronLOCKR-mediated control.

Main Methods:

• Fusion of degronLOCKR to endogenous signaling molecules in yeast to create synthetic feedback loops.
• Evaluation of degronLOCKR-mediated feedback control on a synthetic gene circuit.
• Rational modification of degronLOCKR proteins to tune feedback behavior.

Main Results:

• Successful generation of synthetic negative and positive feedback in the yeast mating pathway.
• Quantification of feedback control capabilities and operational range in a synthetic gene circuit.
• Demonstration of degronLOCKR's ability to rewire complex endogenous pathways and tune feedback.

Conclusions:

• De novo-designed degronLOCKR proteins provide a powerful platform for engineering feedback control in living cells.
• This technology advances synthetic biology by enabling precise regulation of cellular functions.
• The work highlights the potential of de novo protein design for creating tools for biotechnological and therapeutic applications.