A kind of novelmultifunctionalmodules based onzinc(II)-coordinativecyclenhas been developed, which is utilized to modify low molecular weight polyethylenimine (LMWPEI) obtaininghigh-performanceDNAvectors. A series of in vitro experiments were carried out to explore the performance of the module in improving the key process of gene transfection, such as DNA condensation, serum resistance, cellular uptake, and endosomal escape. The results demonstrate that there is a significant synergistic effect between the functional module and PEI2.5k in the process of breaking through the key barriers of gene transfection. The optimal Zn-PCD mediates 160-fold higher gluciferase activity than commercial transfection reagents PEI25k in ADSC stem cells with more than 90% cell viability and achieves excellent transfection efficiency in diverse cell types, for instance, HepG2 cells, 293T cells, and 293F suspension cells.