Novel folate-conjugated biodegradable multipolymeric nanoparticles (NPs) were constructed and evaluated for potential use in gene delivery to human cervical carcinomas Hela cells, which overexpressed folate receptors. Folate-poly(ethylene glycol)-poly(D: , L: -lactic-co-glycolic acid) (PELGA-F) was synthesized and collaborated with poly-L: -lysine (PLL) to form polymer-polycationic peptide-DNA (PPD) NPs. Fluorescein sodium and polylysine-condensed DNA (PD) were encapsulated in both PELGA nanoparticles (PELGA-NPs) and folate modified nanoparticles (PELGA-F-NPs), which were prepared by a modified solvent extraction/evaporation method. Effects of the folate conjugation and PLL introduction on the uptake of NPs was qualified by fluorescent invert microscopy and quantified by spectrofluorometric measurement, while effect on the gene expression was measured by X-gal staining and luciferase assay, both using Hela cells as an in vitro model. Results showed that cellular uptake of NPs was enhanced by folate modification, but had no difference after PLL encapsulation. In transfection tests, increased gene expression also confirmed the different functions of folate and PLL introduction. It is feasible that folate-linked multipolymeric NPs should be an efficient targeted carrier for gene delivery.