Replication-deficient adenovirus gene transfer vectors are very useful for the experimental delivery of genes into cells and are widely used both in vitro and in vivo to determine the effects of transgene expression. Having a broad cell tropism, these vectors allow efficient transduction of many cell types and permit transfer of large amounts of DNA with resulting high expression levels within the target cell. Manganese superoxide dismutase (MnSOD), copper zinc superoxide dismutase (CuZnSOD) and catalase are all known antioxidants whose over-expression can result in amelioration of pathology brought about by an excess of reactive oxygen species within a cell. Their use has been suggested as therapies for many conditions, including cardiovascular disease, arthritis, diabetes, cancer, and damage to central nervous system cells. This chapter describes the methodology commonly used for production of replication-deficient adenovirus vectors encoding MnSOD, CuZnSOD, and catalase.