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A Multilayer Microfluidic Platform for the Conduction of Prolonged Cell-Free Gene Expression

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Synthetic GPCRs for programmable sensing and control of cell behaviour.

Nicholas A Kalogriopoulos¹, Reika Tei¹, Yuqi Yan²

¹Department of Genetics, Stanford University, Stanford, CA, USA.

|December 5, 2024

Summary

Researchers developed programmable antigen-gated G-protein-coupled engineered receptors (PAGERs) that control cellular functions in response to specific antigens. This synthetic biology tool offers modularity and broad applications in therapeutics and research.

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Using Synthetic Biology to Engineer Living Cells That Interface with Programmable Materials

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Published on: March 9, 2017

Area of Science:

Synthetic biology
Molecular engineering
Cellular engineering

Background:

Established synthetic receptors have limitations in detecting soluble antigens and lack integrated drug control.
Chimeric antigen receptors are restricted to immobilized antigens and have a limited output scope.

Purpose of the Study:

To engineer novel synthetic G-protein-coupled receptors (GPCRs) for antigen-dependent control of cellular processes.
To develop a modular platform for programmable antigen-gated G-protein-coupled engineered receptors (PAGERs).

Main Methods:

Engineered GPCR scaffolds with a conditional auto-inhibitory domain fused to nanobody binders.
Achieved modular antigen gating where antigen binding relieves auto-inhibition, enabling drug-induced receptor activation.
Created PAGERs responsive to diverse soluble and cell-surface antigens.

Main Results:

Demonstrated PAGERs driving transgene expression, real-time fluorescence, and endogenous G-protein activation.
Showcased applications including T cell migration, macrophage differentiation, therapeutic antibody secretion, and neuronal activity control.
Successfully generated PAGERs for over a dozen biologically relevant antigens.

Conclusions:

PAGERs provide a versatile and programmable platform for antigen-specific cellular control.
The modular design of PAGERs facilitates broad utility in drug discovery and translational science.
This technology enables precise manipulation of cellular functions through synthetic receptors.