Spatio-Temporal Photoactivation of Cytotoxic Proteins

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.