Intracellular Delivery of Functional Proteins with DNA-Protein Nanogels-Lipids Complex

Area of Science:

• Biotechnology
• Nanomedicine
• Cellular Biology

Background:

• Therapeutic protein delivery faces challenges in intracellular transport.
• Nucleic acid delivery is established but protein delivery lags.
• Functional proteins offer high selectivity and catalytic activity for cellular control.

Purpose of the Study:

• To develop a functionalizable nanoplatform for intracellular protein delivery.
• To demonstrate the platform's ability to deliver active proteins into living cells.
• To assess the platform's potential for biomedical applications.

Main Methods:

• Fabrication of DNA-protein nanogels cross-linked via biotin-streptavidin interactions.
• Loading nanogels with proteins (biotin, streptavidin, or strep-tag).
• Complexation with cationic lipid vectors for cellular delivery.
• Delivery and activity assessment of alkaline phosphatase in mouse melanoma cells.

Main Results:

• Nanogels successfully delivered functional proteins into living cells.
• Delivered alkaline phosphatase retained catalytic activity intracellularly.
• Nanogels are approximately 100 nm in size, carrying ~100 enzyme molecules.
• Effective transfection achieved at low lipid concentrations, indicating low toxicity.

Conclusions:

• The DNA-protein nanogel platform enables effective intracellular delivery of active proteins.
• The system demonstrates low toxicity and high local enzyme concentration.
• This nanoplatform holds significant promise for various biomedical applications.