Multiblock reducible copolypeptides containing histidine-rich and nuclear localization sequences for gene delivery.
Polyplexes sensitive to redox potential gradients represent promising gene delivery vectors. High molecular weight polypeptides containing disulfide bonds in the backbone were synthesized by an oxidative copolymerization of a histidine-rich peptide (HRP) and a nuclear localization sequence (NLS) peptide. The synthetic approach allowed an easy synthesis of reducible copolypeptides (rCPP) with different relative contents of the HRP and NLS sequences. Cytotoxicity and transfection activity of rCPP-based DNA polyplexes were evaluated in vitro. In comparison with control polyethylenimine (PEI), only minimum toxic effects of rCPPs were observed on the metabolic activity and membrane integrity of human endothelial cells. These findings are predominantly ascribed to favorable structural features like lower charge density and higher chain rigidity of the rCPPs compared to PEI and also to a reductive intracellular and plasma membrane degradation. Transfection activity in all tested cell lines increased with increasing content of the HRP sequence in the rCPPs, while no clearly measurable effect of the NLS sequence was found.
Manickam
Devika Soundara
DS
Department of Pharmaceutical Sciences, Wayne State University, Detroit, Michigan 48202, USA.
Oupický
David
D
D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002522:Chlorocebus aethiops; D004247:DNA / Q000737:chemistry / Q000235:genetics; D018014:Gene Transfer Techniques; D006639:Histidine / Q000737:chemistry*; D006801:Humans; D051379:Mice; D019913:Nuclear Localization Signals; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry* / Q000633:toxicity; D014162:Transfection
D019913:Nuclear Localization Signals; D010455:Peptides; D006639:Histidine; D004247:DNA