Engineering of an Optogenetic T Cell Receptor Compatible with Fluorescence-Based Readouts

Area of Science:

• Immunology
• Molecular Biology
• Biotechnology

Background:

• Optogenetics enables precise control over biological signaling pathways.
• T cell receptor (TCR) interactions are crucial for immune responses.
• Modulating TCR activity with external stimuli remains a challenge.

Purpose of the Study:

• To develop an optogenetic system for light-inducible control of T cell receptor (TCR) and ligand interactions.
• To engineer a modified TCR capable of binding to a light-sensitive ligand.
• To establish a system for on-demand manipulation of immune signaling.

Main Methods:

• Engineering of modified T cell receptors (TCRs) fused with phytochrome B-interacting factors (PIF^S).
• Testing various PIF^S-fused TCR constructs for surface expression and ligand binding.
• Utilizing the plant photoreceptor phytochrome B (PhyB) as a light-activated ligand.
• Combining the engineered TCR with a GFP reporter system driven by NF-AT.

Main Results:

• The SNAP-PIF^S-TCR construct demonstrated successful surface expression and binding to PhyB.
• Light-induced interaction between SNAP-PIF^S-TCR and PhyB elicited downstream activation signals.
• The system was compatible with a reporter assay for monitoring T cell activation via GFP expression.

Conclusions:

• A novel optogenetic system was successfully developed to control T cell receptor signaling with light.
• The SNAP-PIF^S-TCR provides a light-switchable module for modulating immune cell activation.
• This technology offers potential for precise spatiotemporal control in immunotherapy and immunological research.