Abstract
A novel gene delivery system based on the enhanced release of plasmid DNA from polyplexes in response to intracellular proprotein convertase (PC), furin, that cleaves the carboxyl side of Arg-X-Arg/Lys-Arg, was successfully established. Polypeptides that have this recognition sequence were synthesized by the Fmoc solid phase method and evaluated as gene vectors. The fragmented polypeptides were found to lose the abilities to form polyplexes with plasmid DNA. When the Fur-polypeptide/plasmid DNA polyplexes were treated with furin in vitro, their disassembly and the release of plasmid DNA was observed in agarose gel electrophoresis analysis. The Fur-polypeptide showed higher transgene expression in COS-1 cells, while the control polypeptides which do not have the recognition sites did not. These results indicate that the cationic nature of the polypeptides is not sufficient for gene carrier and this intracellular signal-responsive gene transfer system is the useful strategy to enhance the transcription efficiency of the delivered plasmid DNA.
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