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The efficacy of gene therapy using nonviral vector based on polyplex micelle has been studied against corneal neovascularization in mice.A block copolymer, poly(ethylene glycol) (PEG)-block-polycation carrying ethylenediamine units in the side chain (PEG-b-P[Asp(DET)]), was prepared. PEG-b-P[Asp(DET)] formed a polyplex micelle through the polyion complex formation with plasmid DNA. To evaluate in vivo gene transfer efficiency, PEG-b-P[Asp(DET)] micelle was injected into the subconjunctival space of mice, and the expression of the reporter gene was assessed. Furthermore, mouse corneal neovascularization models were treated with the PEG-b-P[Asp(DET)] polyplex micelle containing expression plasmid vector of soluble vascular endothelial growth factor receptor 1 (sflt-1).Subconjunctival injection of the PEG-b-P[Asp(DET)] polyplex micelle containing a reporter gene showed prolonged gene expression with low cytotoxicity. Also, gene transfer into subconjunctival space by the polyplex micelle containing sflt-1 plasmid showed significant inhibition of corneal neovascularization in mice.Nonviral gene therapy using PEG-b-P[Asp(DET)] polyplex micelle may have potential for safe and effective therapeutic treatment of corneal neovascularization.