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Updated: Sep 27, 2025
Spatial and Temporal Control of T Cell Activation Using a Photoactivatable Agonist
Published on: April 25, 2018
Spatio-Temporal Photoactivation of Cytotoxic Proteins
Raquel Cruz-Samperio1, Robert J Mart1, Louis Y P Luk1
1School of Chemistry, Cardiff University, Main Building, Park Place, Cardiff, CF10 3AT, U.K.
Abstract:
Protein therapeutics offer exquisite selectivity in targeting cellular processes and behaviors, but are rarely used against non-cell surface targets due to their poor cellular uptake. While cell-penetrating peptides can be used to deliver recombinant proteins to the cytosol, it is generally difficult to selectively deliver active proteins to target cells. Here, we report a recombinantly produced, intracellular protein delivery and targeting platform that uses a photocaged intein to regulate the spatio-temporal activation of protein activity in selected cells upon irradiation with light. The platform was successfully demonstrated for two cytotoxic proteins to selectively kill cancer cells after photoactivation of intein splicing. This platform can generically be applied to any protein whose activity can be disrupted by a fused intein, allowing it to underpin a wide variety of future protein therapeutics.
Insights
Researchers developed a novel protein delivery platform using a light-activated intein for precise, spatio-temporal control of protein therapeutics. This technology enables selective activation of intracellular protein activity, enhancing targeted cancer cell killing.
Area of Science:
- Biotechnology
- Molecular Biology
- Drug Delivery
Background:
- Protein therapeutics offer high selectivity but face challenges in intracellular delivery and targeting.
- Poor cellular uptake limits the application of protein drugs to non-cell surface targets.
- Existing methods for intracellular protein delivery often lack precise spatial and temporal control.
Purpose of the Study:
- To develop a novel platform for the controlled intracellular delivery and activation of protein therapeutics.
- To enable spatio-temporal regulation of protein activity within selected cells using light.
- To demonstrate the platform's efficacy in selectively targeting and killing cancer cells.
Main Methods:
- Engineered a recombinantly produced platform utilizing a photocaged intein to control protein activity.
- Developed a system for light-induced activation of intein splicing, releasing active protein intracellularly.
- Tested the platform with two cytotoxic proteins to induce cancer cell death upon photoactivation.
Main Results:
- Successfully demonstrated spatio-temporal control over protein activity in selected cells using light.
- Achieved selective killing of cancer cells through photoactivated intein splicing and release of cytotoxic proteins.
- Validated the generic applicability of the platform for various protein therapeutics.
Conclusions:
- The photocaged intein platform offers a versatile strategy for intracellular protein delivery and targeted activation.
- This technology overcomes limitations of cellular uptake and enhances selectivity for protein therapeutics.
- The platform holds significant potential for developing next-generation protein-based drugs for various diseases.
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