A number of methods have been developed for gene delivery including recombinant viral vectors, synthetic polymers, peptides, lipids and targeted vesicles. Recombinant proteins, with their inherent biocompatibility, versatility for incorporating targeting function, low toxicity, and facile preparation, have shown great advantages as vehicles for non-viral gene delivery. In this study, a recombinant protein was developed for delivering DNA oligonlucleotides to PC12 cells. This recombinant protein (p50-GFP-TAT) consists of a DNA binding domain (p50), a cell penetrating domain (TAT), and a fluorescent protein domain (GFP). Two other recombinant proteins, p50-GFP and GFP-TAT, were also prepared as controls. The efficiency of this protein vehicle was evaluated by delivering fluorescently-labeled dsDNA molecules of 30bp and 294bp lengths at varying molar ratios of protein to DNA in a physiological medium containing 17.5% serum. The results show that the three recombinant protein constructs can transfect PC12 cells in a dose dependent pattern but p50-GFP-TAT exhibits the highest transfection efficiency compared to p50-GFP and GFP-TAT. However, among the three proteins, only p50-GFP-TAT can deliver DNA molecules to the PC-12 cells, which indicates both DNA binding domain (p50) and cell penetrating domain (TAT) are necessary for the successful delivery of DNA. The optimal molar ratios of protein to DNA are 11.2 for delivery of the 30bp DNA oligonucleotide and 560 for delivery of 294bp DNA oligonucleotide. Our study has also shown that TAT exhibits inherent transfection specificity to different cell types and even the same cell type under varying conditions. For example the GFP-TAT peptide preferentially penetrates serum-stimulated astrocytes over quiescent astrocytes and neurons grown in a serum-free environment, which indicates a potential application of the construct in targeting activated astrocytes for the treatment of brain injuries. Pilot studies exploring the efficiency of the recombinant protein containing both p50 and TAT domain for the delivery of siRNA will also be presented. Overall, this study will demonstrate the potential of designed recombinant protein constructs as non-viral gene therapy vehicles for the delivery of various DNA and RNA cargoes.