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In Vitro Gene Delivery in Retinal Pigment Epithelium Cells by Plasmid DNA-Wrapped Gold Nanoparticles

1
Department of Zoology, University of Oxford, Oxford OX1 3PS, UK
2
Departament de Genètica, Microbiologia i Estadística, Universitat de Barcelona, 08028 Barcelona, Spain
3
CIBERER, ISCIII, Universitat de Barcelona, 08028 Barcelona, Spain
4
Institute of Biomedicine (IBUB-IRSJD), Universitat de Barcelona, 08028 Barcelona, Spain
*
Authors to whom correspondence should be addressed.
Genes 2019, 10(4), 289; https://doi.org/10.3390/genes10040289
Received: 19 March 2019 / Revised: 5 April 2019 / Accepted: 8 April 2019 / Published: 9 April 2019
(This article belongs to the Special Issue Molecular Therapies for Inherited Retinal Diseases)
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Abstract

Many rare diseases course with affectation of neurosensory organs. Among them, the neuroepithelial retina is very vulnerable due to constant light/oxidative stress, but it is also the most accessible and amenable to gene manipulation. Currently, gene addition therapies targeting retinal tissue (either photoreceptors or the retinal pigment epithelium), as a therapy for inherited retinal dystrophies, use adeno-associated virus (AAV)-based approaches. However, efficiency and safety of therapeutic strategies are relevant issues that are not always resolved in virus-based gene delivery and alternative methodologies should be explored. Based on our experience, we are currently assessing the novel physical properties at the nanoscale of inorganic gold nanoparticles for delivering genes to the retinal pigment epithelium (RPE) as a safe and efficient alternative approach. In this work, we present our preliminary results using DNA-wrapped gold nanoparticles (DNA-gold NPs) for successful in vitro gene delivery on human retinal pigment epithelium cell cultures, as a proof-of-principle to assess its feasibility for retina in vivo gene delivery. Our results show faster expression of a reporter gene in cells transfected with DNA-gold NPs compared to DNA-liposome complexes. Furthermore, we show that the DNA-gold NPs follow different uptake, internalization and intracellular vesicle trafficking routes compared to pristine NPs. View Full-Text
Keywords: gene therapy; gold nanoparticles; DNA-wrapped gold nanoparticles; ARPE-19 cells; retinal pigment epithelium; clathrin-coated vesicles; endosomal trafficking gene therapy; gold nanoparticles; DNA-wrapped gold nanoparticles; ARPE-19 cells; retinal pigment epithelium; clathrin-coated vesicles; endosomal trafficking
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Trigueros, S.; B. Domènech, E.; Toulis, V.; Marfany, G. In Vitro Gene Delivery in Retinal Pigment Epithelium Cells by Plasmid DNA-Wrapped Gold Nanoparticles. Genes 2019, 10, 289.

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