Journal Article

FZJ-2025-03167

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Effect of degeneration stage on non-viral tissue transfection of rd10 retina ex vivo

Zhang, J. ; Bing, Z. ; Marie, C. ; Izsvák, Z. ; Müller, F.FZJ* ; Thumann, G. ; Walter, P.FZJ* ; Johnen, S. (Corresponding author)

2025
Nature Publ. Group New York, NY

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Please use a persistent id in citations: doi:10.1016/j.omtn.2025.102616  doi:10.34734/FZJ-2025-03167

Abstract: Gene therapy has great potential for the treatment of inherited retinal diseases, as evidenced by the progress and ongoing research. Using the Sleeping Beauty (SB) transposon system, we developed a non-viral gene delivery system for electroporation-based transfection of rd10 retinas ex vivo. SB100X transposase and Venus transposon plasmids were transfected at a ratio of 1:16 into rd10 retinas of different ages and corresponding wild-type (WT) controls. Transfection efficiency was assessed by fluorescence microscopy and transfected cells were identified by immunohistochemistry. Retinal integrity and cell viability were assessed by FITC-dextran electroporation, histology, and apoptosis assay. The highest transfection efficiency was observed in degenerated stages P61 and older, with Müller cells being the only transfected cell type. A 31% reduction in transposon plasmid size resulted in a 1.5-fold increase in transfection efficiency. Integrity and morphology of degenerated retinas were preserved after electroporation-based plasmid transfer; the number of apoptotic cells in the inner nuclear layer (INL) was reduced by half compared to WT controls. We demonstrated that electroporation-based delivery of the SB transposon system resulted in efficient transfection of degenerated retinas. Our results are an important first step toward the combined use of retinal prostheses and gene therapy to improve the treatment of inherited retinal dystrophies.

Note: This work was supported by the Confocal Microscope Facility and the Immunohistochemistry Facility, core facilities of the Interdisciplinary Center for Clinical Research (IZKF) Aachen within the Faculty of Medicine at RWTH Aachen University. Thethors thank Anne Freialdenhoven, Antje Schiefer, and Alicia Hühnlein (Department of Ophthalmology, University Hospital RWTH Aachen) for excellent technicaltance. This work was funded by the China Scholarship Council (no. 202208080251) and is part of a graduate school funded by the Deutsche Forschungsgemeinschaft DFG under GRK 2610/01. The results shown here are part of the doctoral thesis of J.Z.

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Contributing Institute(s):

1. Molekular- und Zellphysiologie (IBI-1)
2. Katalytische Grenzflächen (INW-1)

Research Program(s):

1. 5244 - Information Processing in Neuronal Networks (POF4-524) (POF4-524)
2. GRK 2610 - GRK 2610: Innovative Schnittstellen zur Retina für optimiertes künstliches Sehen - InnoRetVision (424556709) (424556709)

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