DNA Vaccination

Jeffrey B. Ulmer1, Rino Rappuoli1

1 Chiron Corporation, Emeryville, California
Publication Name:  Current Protocols in Human Genetics
Unit Number:  Unit 13.2
DOI:  10.1002/0471142905.hg1302s37
Online Posting Date:  August, 2003
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Abstract

This unit details some of the key methods for setting up and testing DNA vaccines in animal models. The basic procedures are discussed, as well as alternative methods that have been developed over the past several years. The gives step‐by‐step instructions for administering the DNA vaccine via intramuscular injection of the quadriceps muscle, while an alternate procedure details injection of the anterior tibialis.

     
 
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Table of Contents

  • Basic Protocol 1: DNAVaccine Administration by Intramuscular Injection of the Quadricpeps Muscles in the Mouse
  • Alternate Protocol 1: DNA Vaccine Administration by Intramuscular Injection of the Anterior Tibialis in the Mouse
  • Reagents and Solutions
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: DNAVaccine Administration by Intramuscular Injection of the Quadricpeps Muscles in the Mouse

  Materials
  • Mouse (C57BL/6 and BALB/c strains are commonly used)
  • recipeAnesthetic solution (see recipe)
  • recipeDNA vaccine solution (see recipe; also see )
  • 0.3‐ and 0.1‐ml syringes
  • 26‐G and 28‐G, 0.5‐in. needles
  • Electric clippers
  • Cotton balls or gauze pads
  • Additional reagents and equipment for handling and injection of mice ( appendix 3L)

Alternate Protocol 1: DNA Vaccine Administration by Intramuscular Injection of the Anterior Tibialis in the Mouse

  • Polyethylene tubing, i.d. 0.38‐mm
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Figures

  •   FigureFigure 13.2.1 Anatomy of mouse by muscles.
  •   FigureFigure 13.2.2 A typical DNA vaccine vector contains several important elements, including the promoter, transcription terminator, bacterial origin of replication, antibiotic resistance gene, and antigen gene. Conventional DNA vaccines encode simply the antigen gene, whereas alphavirus‐based DNA vaccines encode an alphavirus RNA replicon with a subgenomic (SG) promoter driving expression of the antigen.
  •   FigureFigure 13.2.3 Steps in generating a DNA vaccine for animal testing. Potential problems can arise due to defective construction, contamination during purification, and inherently poor immunogenicity of the vaccine; potential solutions to these problems are shown as well (also see ).

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Literature Cited

Literature Cited
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