Addition of free poloxamer 407 to a new gene vector P407-PEI-K12 solution forms a sustained-release in situ hypergel that enhances cell transfection and extends gene expression.

Abstract

To address the concern around the efficiency/cytotoxicity ratio and the tumor-targeting effects of polyethylenimine (PEI), is a non-viral gene vector used for the delivery of the cancer therapy gene, poloxamer 407 (P407)-PEI-K12, was synthesized by cross-linking low-molecular weight PEI with P407 and further coupling a bifunctional peptide, K12, which is comprised of the tumor-targeting peptide tLyP-1 and the nuclear localization sequence. Furthermore, the addition of free P407 into the polymer/DNA complex solution produced a temperature-sensitive in situ gel-P407/P407-PEI-K12/DNA complex, which improved the effects of sustained-release gene delivery and transfection efficiency. The specificity, cytotoxicity and gene transfection efficiency of P407-PEI-K12 was investigated in Hela cells in vitro. The polymer efficiently prevented the degradation of plasmid DNA by DNase I and had a marked ability for serum tolerance. Agarose gel electrophoresis revealed that plasmid DNA was efficiently condensed and protected. The higher transfection efficiency of P407-PEI-K12h (the molar ratio of P407-PEI and K12 is 1:10) was achieved with a polymer and plasmid DNA ratio (w/w) of 20:1. The ability of free P407 to promote the transfection of the polymer/DNA complex was high (0.09%). The half-life of the P407/P407-PEI-K12-h/DNA gel complex was 228 min, and the transfection efficiency of the P407/P407-PEI-K12-h/DNA complex was markedly higher compared to that of the P407-PEI-K12-h/DNA complex at various release times.