[Construction and expression analysis of micro-linear vector as a new general gene therapy vector]

Area of Science:

• Molecular Biology
• Gene Therapy
• Biotechnology

Context:

• Gene therapy faces challenges in balancing effective gene delivery with patient safety.
• Traditional viral and plasmid vectors can elicit adverse immunological responses due to non-therapeutic gene expression.

Purpose:

• To develop a novel, safe, and efficient gene therapy vector system named the Micro-Linear Vector.
• To engineer a vector that minimizes non-therapeutic DNA and protects the therapeutic gene cassette from degradation.

Summary:

• The Micro-Linear Vector features a unique cap protecting the gene expression cassette (promoter, enhancer, objective gene, RNA-stabilizing sequence) from exnuclease degradation.
• Utilizing GFP as a reporter gene, the Micro-Linear Vector demonstrated superior transfection efficiency and reduced toxicity compared to the pEGFP-N3 plasmid in 293, 3T3, CNE2, and B95-8 cell lines.
• Experimental validation involved fluorescence microscopy and flow cytometry to assess transfection rates and cellular responses.

Impact:

• The Micro-Linear Vector presents a promising alternative to conventional vectors, potentially enhancing the safety and efficacy of gene therapy applications.
• This innovation could lead to reduced side effects and improved patient outcomes in various genetic disorder treatments.