Designed protein logic to target cells with precise combinations of surface antigens

Area of Science:

• Biochemistry
• Synthetic Biology
• Immunology

Background:

• Mammalian cell identification is complex due to the lack of unique surface markers.
• Targeting specific cell populations requires methods beyond single-marker recognition.

Purpose of the Study:

• To design novel protein-based switches capable of performing Boolean logic operations (AND, OR, NOT).
• To enable precise, transcription-independent cellular responses at single-cell resolution within complex biological systems.
• To demonstrate the application of these switches in redirecting T cell specificity for targeted cancer therapy.

Main Methods:

• Design and engineering of colocalization-dependent protein switches (Co-LOCKR).
• Implementation of Co-LOCKR systems to perform AND, OR, and NOT logic gates.
• Application of Co-LOCKR for T cell redirection against tumor cells with specific surface antigen combinations.

Main Results:

• Co-LOCKR switches successfully executed AND, OR, and NOT Boolean logic operations.
• Engineered AND gates enabled T cell targeting of tumor cells expressing dual antigens while avoiding single-antigen cells.
• Three-input switches integrated NOT or OR logic to refine cellular targeting based on additional antigen presence.

Conclusions:

• De novo designed proteins can perform computational logic on cellular surfaces.
• Co-LOCKR technology offers a versatile platform for precise cell targeting and therapeutic applications.
• This approach allows for sophisticated integration of multiple binding events into a single, functional output.