Engineering T7 bacteriophage as a potential DNA vaccine targeting delivery vector

Area of Science:

• Biotechnology
• Molecular Biology
• Virology

Background:

• Bacteriophage have been utilized for DNA delivery.
• Surface display of cell-penetrating peptides, such as the Tat peptide from HIV-1, enhances DNA transfer.
• Previous studies have not explored T7 phage for this application.

Purpose of the Study:

• To engineer T7 phage as a DNA delivery vector.
• To facilitate cellular internalization of genetic material using T7 phage.
• To evaluate T7 phage displaying Tat peptide for DNA vaccine applications.

Main Methods:

• Constructed recombinant T7 phages displaying the Tat peptide on their surface.
• Incorporated a eukaryotic expression box (EEB) into the T7 phage genome (T7-EEB-Tat).
• Assessed the packaging of foreign gene insertions into infective progeny phage particles.

Main Results:

• Successfully packaged foreign gene insertions into infective T7 phage particles.
• Mammalian cells exposed to T7-EEB-Tat showed significantly higher marker gene expression compared to controls.
• Control cells infected with wild-type T7 phage lacking Tat peptides showed lower expression levels.

Conclusions:

• T7 phage can be engineered as a functional DNA delivery vector.
• Surface display of Tat peptide enhances cellular uptake and gene expression.
• T7 phage holds potential as an effective delivery vector for DNA vaccines.