Optimization of human papillomavirus-based pseudovirus techniques for efficient gene transfer.

Area of Science:

• Molecular biology
• Virology
• Gene delivery technologies

Background:

• Human papillomavirus (HPV) capsid proteins (L1 and L2) self-assemble into virus-like particles.
• These capsids can package the viral genome or non-viral DNA, presenting a potential tool for gene delivery.
• Current methods for using HPV capsids for non-viral DNA delivery require optimization.

Purpose of the Study:

• To optimize existing protocols and introduce new methods for utilizing HPV capsids to deliver non-viral DNA.
• To provide an alternative to traditional DNA transfection methods.
• To enhance the efficiency of HPV capsid-mediated gene delivery in various cell types.

Main Methods:

• Optimization of HPV capsid assembly and DNA packaging protocols.
• Development of novel suspension-based techniques for cell infection.
• Utilizing keratinocyte-derived extracellular matrices to enhance infection efficiency.

Main Results:

• Demonstrated efficient packaging of non-viral DNA by HPV capsids.
• Achieved enhanced infection efficiency in keratinocytes, hepatocytes, and neuronal cells using optimized methods.
• Successfully developed a suspension-based infection technique applicable to diverse cell types.

Conclusions:

• HPV capsids represent a viable and efficient alternative to DNA transfection for delivering non-viral DNA.
• Optimized protocols and novel techniques significantly improve gene delivery efficiency across multiple cell types.
• The developed methods offer a versatile platform for studying proteins of interest in various cellular contexts.