DNA vaccines could offer a solution to a number of problems faced by the poultry industry; they are relatively easy to manufacture, stable, potentially easy to administer, can overcome neonatal tolerance and the deleterious effects of maternal antibody, and do not cause disease pathology. Combined with this, in ovo vaccination offers the advantage of reduced labor costs, mass administration and the induction of an earlier immune response. Together, this list of advantages is impressive. However, this combined technology is still in its infancy and requires many improvements. The potential of CpG motifs, DNA vaccines and in ovo vaccination, however, can be observed by the increasing number of recent reports investigating their application in challenge experiments. CpG motifs have been demonstrated to be stimulatory both in vitro and in vivo. In addition, DNA vaccines have been successfully delivered via the in ovo route, albeit not yet through the amniotic fluid. Lastly, a recent report has demonstrated that a DNA vaccine against infectious bronchitis virus administered via in ovo vaccination, followed by live virus boost, can slightly improve on the protective effect induced by the live virus alone. Therefore, DNA vaccination via the in ovo route is promising and offers potential as a poultry vaccine, however, efficacy needs to be improved and the costs of production reduced before it is likely to be beneficial to the poultry industry in the long term.