A novel strategy to improve the transfection efficiency as well as lower the cytotoxicity for PDMAEMA polycationic vectors has been developed. The cytotoxicity decreases and the transfection efficiency increases significantly through the incorporation of a hydrophobic hyperbranched PEHO core into PDMAEMA. The effectiveness of the hyperbranched architecture for the gene transfection was further proved by comparing the PEHO-star-PDMAEMAs vector with a linear, molecular weight-analogous PEHO core as the control.