Rapid transport of plasmid DNA into the nucleolus via actin depolymerization using the HVJ envelope vector

Area of Science:

• Molecular Biology
• Cell Biology
• Gene Therapy

Background:

• Nuclear transport of therapeutic genes is crucial for human gene therapy.
• Non-viral vectors improve exogenous DNA delivery but face challenges in nuclear transport.
• Efficient nuclear entry of plasmid DNA remains a key area for research.

Purpose of the Study:

• To investigate the nuclear translocation efficiency of plasmid DNA delivered by different non-viral vectors.
• To determine the effect of actin depolymerization on the nuclear transport of plasmid DNA.

Main Methods:

• Comparison of nuclear entry of Cy3-labeled plasmid DNA (Cy3-pDNA) using hemagglutinating virus of Japan envelope (HVJ-E) vector, Lipofectamine, and microinjection.
• Assessment of the impact of actin depolymerization on Cy3-pDNA nuclear transport.

Main Results:

• HVJ-E vector mediated rapid nuclear and nucleolar entry of Cy3-pDNA within 30 minutes.
• Cationic liposomes resulted in cytoplasmic retention of Cy3-pDNA.
• HVJ-E treatment induced transient actin depolymerization, accelerating nucleolar entry of microinjected DNA.

Conclusions:

• Plasmid DNA transport to the nucleolus is accelerated by actin filament depolymerization.
• The HVJ-E vector enhances DNA transport to the nucleolus through actin depolymerization, offering a promising strategy for gene therapy.