AIM: To design and develop a novel target-specific DNA-delivery system using hyaluronic acid (HA)-polyamidoamine (PAMAM) conjugates (P-HA). MATERIALS & METHODS: The coupling of HA to the PAMAM dendrimer was analyzed by (1)H-NMR and elemental analysis (CHN). Their properties were characterized in terms of size and zeta-potential and evaluated for in vitro and in vivo transfection efficiency. RESULTS: The designed covalent HA-dendriplexes enhanced gene transfection of pCMV-Luc reporter gene in overexpressing CD44-receptor cancer cells. They were also more efficient in transfecting MDA-MB231 cells than conventional PEI-polyplexes. The cytotoxicity of the covalent HA-dendriplexes was lower than when using conventional polyethylenimine-polyplexes. In vivo studies showed that these targeted complexes were also efficient for delivering pCMVLuc in different organs of healthy mice, as well as in tumors of C57BL/6 animals. CONCLUSIONS: The HA-dendriplexes developed in this work may offer an advantageous alternative to conventional cationic polymer-based formulations for DNA delivery into cancer cells in an efficient and safe manner.