BACKGROUND: We have previously shown that a novel synthetic peptide for ocular delivery (POD) can efficiently compact DNA and deliver it to cells in vitro. This observation prompted us to develop use of POD as a nonviral vector in vivo. METHODS: POD peptide was modified using poly(ethylene) glycol (PEG-POD) and used to compact DNA into nanoparticles that were then analysed using electron microscopy, dynamic light scattering, and fluorescent labeling. Transfection efficiency and localization were determined 48 h post-injection into the subretinal space of the mouse eye using luciferase and LacZ, respectively. Efficiency of ocular transfection was compared to two other PEGylated peptides: PEG-TAT and PEG-CK30. RESULTS: PEG-POD can compact DNA and form discrete nanoparticles of approximately 136 nm that can penetrate and transduce the retinal pigment epithelium (RPE) in vivo. PEG-POD significantly increased expression of plasmid DNA by 215-fold, PEG-TAT by 56.52-fold, and PEG-CK30 by 24.73-fold relative to DNA injected alone. In all cases beta-galactosidase was observed primarily in the RPE layer after subretinal injection. Electrophysiological analyses of PEG-POD transduced retina indicates an absence of PEG-POD-mediated toxicity. PEG-POD can protect plasmid DNA from DNaseI digestion, resulting in significant transfection of the lung after intravenous injection in mice. CONCLUSIONS: PEG-POD was found to significantly increase gene delivery relative to both DNA alone and other pegylated peptides. These findings highlight the use of pegylated peptides, and specifically PEG-POD, as novel gene delivery vectors. Copyright 2009 John Wiley & Sons, Ltd.
BACKGROUND: We have previously shown that a novel synthetic peptide for ocular delivery (POD) can efficiently compact DNA and deliver it to cells in vitro. This observation prompted us to develop use of POD as a nonviral vector in vivo. METHODS:POD peptide was modified using poly(ethylene) glycol (PEG-POD) and used to compact DNA into nanoparticles that were then analysed using electron microscopy, dynamic light scattering, and fluorescent labeling. Transfection efficiency and localization were determined 48 h post-injection into the subretinal space of the mouse eye using luciferase and LacZ, respectively. Efficiency of ocular transfection was compared to two other PEGylated peptides: PEG-TAT and PEG-CK30. RESULTS:PEG-POD can compact DNA and form discrete nanoparticles of approximately 136 nm that can penetrate and transduce the retinal pigment epithelium (RPE) in vivo. PEG-POD significantly increased expression of plasmid DNA by 215-fold, PEG-TAT by 56.52-fold, and PEG-CK30 by 24.73-fold relative to DNA injected alone. In all cases beta-galactosidase was observed primarily in the RPE layer after subretinal injection. Electrophysiological analyses of PEG-POD transduced retina indicates an absence of PEG-POD-mediated toxicity. PEG-POD can protect plasmid DNA from DNaseI digestion, resulting in significant transfection of the lung after intravenous injection in mice. CONCLUSIONS:PEG-POD was found to significantly increase gene delivery relative to both DNA alone and other pegylated peptides. These findings highlight the use of pegylated peptides, and specifically PEG-POD, as novel gene delivery vectors. Copyright 2009 John Wiley & Sons, Ltd.
Authors: Ge Liu; DeShan Li; Murali K Pasumarthy; Tomasz H Kowalczyk; Christopher R Gedeon; Susannah L Hyatt; Jennifer M Payne; Timothy J Miller; Peter Brunovskis; Tamara L Fink; Osman Muhammad; Robert C Moen; Richard W Hanson; Mark J Cooper Journal: J Biol Chem Date: 2003-06-14 Impact factor: 5.157
Authors: M Neuner-Jehle; L V Berghe; S Bonnel; Y Uteza; F Benmeziane; J S Rouillot; D Marchant; A Kobetz; J L Dufier; M Menasche; M Abitbol Journal: Hum Gene Ther Date: 2000-09-01 Impact factor: 5.695
Authors: Assem-Galal Ziady; Christopher R Gedeon; Timothy Miller; William Quan; Jennifer M Payne; Susannah L Hyatt; Tamara L Fink; Osman Muhammad; Sharon Oette; Tomasz Kowalczyk; Murali K Pasumarthy; Robert C Moen; Mark J Cooper; Pamela B Davis Journal: Mol Ther Date: 2003-12 Impact factor: 11.454
Authors: Assem-Galal Ziady; Christopher R Gedeon; Osman Muhammad; Virginia Stillwell; Sharon M Oette; Tamara L Fink; Will Quan; Tomasz H Kowalczyk; Susannah L Hyatt; Jennifer Payne; Angela Peischl; J E Seng; Robert C Moen; Mark J Cooper; Pamela B Davis Journal: Mol Ther Date: 2003-12 Impact factor: 11.454
Authors: Irina A Ignatovich; Ella B Dizhe; Anna V Pavlotskaya; Boris N Akifiev; Sergey V Burov; Sergey V Orlov; Andrej P Perevozchikov Journal: J Biol Chem Date: 2003-07-25 Impact factor: 5.157