Particle-mediated DNA vaccines employ a physical, intracellular delivery device to achieve the deposition of plasmid DNA-based expression vectors directly into the interior of cells of the skin. The resultant bolus of transient antigen expression in keratinocytes and trafficking dendritic cells results in the induction of humoral and cellular immune responses in various animal models and humans, mimicking characteristics of live or live-vectored vaccines. Ultimately, DNA vaccine success in the clinic will depend on both the successful intracellular delivery of a plasmid vector and an immunostimulator or adjuvant to maximise humoral and cellular immune responses to the encoded antigen(s). To this end, recent DNA vaccine clinical trials are confirming the importance of an intracellular delivery system, while preclinical studies in animal models are demonstrating the feasibility of augmenting responses through the use of DNA-encoded immunostimulators. Particle-mediated DNA vaccines represent a promising tool for developing candidate vaccines against some of the more difficult infectious, parasitic and oncologic disease targets.