Boolean logic-gated protein presentation through autonomously compiled molecular topology.

Area of Science:

• Biomaterials Science
• Chemical Biology
• Synthetic Biology

Background:

• Stimulus-responsive materials are crucial for advanced applications like biosensing and drug delivery.
• Existing methods for creating complex responsive materials often involve difficult and inefficient multi-step organic syntheses.
• Controlled molecular topology is key to designing materials that respond to specific input combinations.

Purpose of the Study:

• To develop a scalable and efficient method for synthesizing topologically specified protein cargos for biomaterials.
• To enable user-programmable Boolean logic for conditional release of cargo from biomaterials.
• To demonstrate the versatility of this approach for various applications including multiplexed delivery and cellular localization.

Main Methods:

• Utilized recombinant expression and chemical biology tools to create topologically specified protein cargos.
• Integrated spontaneous intramolecular ligations for autonomous construct topology compilation during expression.
• Employed orthogonal protease actuators to achieve user-programmable Boolean logic (AND, OR, YES gates).

Main Results:

• Successfully synthesized advanced Boolean logic operators through a direct and scalable expression process.
• Demonstrated all 17 possible logic outputs for protein release based on input combinations of three orthogonal protease actuators.
• Achieved multiplexed delivery of three distinct biomacromolecules from hydrogels and conditional cargo liberation based on five inputs.

Conclusions:

• This novel framework enables the scalable and direct synthesis of advanced, topologically defined protein cargos.
• The developed system offers precise, logic-gate-controlled release of biomolecules from materials.
• The approach has significant potential for applications in advanced drug delivery, biosensing, and cellular engineering.