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Suitable Promoter for DNA Vaccination Using a pDNA Ternary Complex.

Tomoaki Kurosaki¹, Hiroki Nakamura¹, Hitoshi Sasaki²

¹Graduate School of Biomedical Sciences, Nagasaki University, 1-7-1 Sakamoto, Nagasaki 852-8588, Japan.

|May 25, 2024

Summary

The chicken beta-actin hybrid (CBh) promoter enhances gene expression and immune response in plasmid DNA (pDNA) ternary complexes, making it ideal for DNA vaccination.

Keywords:

gene delivery pDNA polyethylenimine promoters γ-polyglutamic acid

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Area of Science:

Biotechnology
Molecular Biology
Immunology

Background:

Plasmid DNA (pDNA) ternary complexes are investigated for gene delivery and vaccination.
Promoter selection significantly impacts gene expression and immune responses.

Purpose of the Study:

To evaluate the influence of different promoters on pDNA ternary complex transfection and immune induction.
To identify the most effective promoter for DNA vaccination applications.

Main Methods:

Constructed model pDNAs with Simian Virus 40 (SV40), Eukaryotic Elongation Factor 1 Alpha (EF1), Cytomegalovirus (CMV), and Chicken Beta-Actin Hybrid (CBh) promoters.
Formulated pDNA ternary complexes with consistent particle size and zeta potential.
Assessed gene expression in cells and spleen after intravenous administration.
Measured anti-ovalbumin (OVA) immunoglobulin G (IgG) induction post-immunization.

Main Results:

pDNA ternary complexes with EF1, CMV, and CBh promoters demonstrated high gene expression.
The SV40 promoter resulted in low gene expression.
Only the pDNA ternary complex with the CBh promoter induced significant anti-OVA IgG antibody production.

Conclusions:

The CBh promoter shows significant potential for enhancing DNA vaccination efficacy.
Promoter choice is critical for optimizing gene expression and immune responses in pDNA ternary complex-based vaccines.