Effect of degeneration stage on non-viral tissue transfection of rd10 retina ex vivo
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...
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Elsevier
2025-09-01
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| Series: | Molecular Therapy: Nucleic Acids |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253125001702 |
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| author | Jiao Zhang Ziyu Bing Corinne Marie Zsuzsanna Izsvák Frank Müller Gabriele Thumann Peter Walter Sandra Johnen |
| author_facet | Jiao Zhang Ziyu Bing Corinne Marie Zsuzsanna Izsvák Frank Müller Gabriele Thumann Peter Walter Sandra Johnen |
| author_sort | Jiao Zhang |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-e040a2d7ac014fbeb0d850ab15095bbb |
| institution | Kabale University |
| issn | 2162-2531 |
| language | English |
| publishDate | 2025-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj-art-e040a2d7ac014fbeb0d850ab15095bbb2025-08-20T03:50:58ZengElsevierMolecular Therapy: Nucleic Acids2162-25312025-09-0136310261610.1016/j.omtn.2025.102616Effect of degeneration stage on non-viral tissue transfection of rd10 retina ex vivoJiao Zhang0Ziyu Bing1Corinne Marie2Zsuzsanna Izsvák3Frank Müller4Gabriele Thumann5Peter Walter6Sandra Johnen7Department of Ophthalmology, Uniklinik RWTH Aachen, 52074 Aachen, GermanyDepartment of Ophthalmology, Uniklinik RWTH Aachen, 52074 Aachen, GermanyUniversité Paris Cité, CNRS, Inserm, UTCBS, 75006 Paris, France; Chimie ParisTech, Université PSL, 75005 Paris, FranceMax Delbrück Center for Molecular Medicine in the Helmholtz Association, 13125 Berlin, GermanyInstitute of Biological Information Processing, Molecular and Cellular Physiology, IBI-1, Forschungszentrum Jülich GmbH, 52428 Jülich, GermanyDepartment of Ophthalmology, University Hospitals of Geneva, 1205 Geneva, Switzerland; Experimental Ophthalmology, University of Geneva, 1205 Geneva, SwitzerlandDepartment of Ophthalmology, Uniklinik RWTH Aachen, 52074 Aachen, GermanyDepartment of Ophthalmology, Uniklinik RWTH Aachen, 52074 Aachen, Germany; Corresponding author: Sandra Johnen, Department of Ophthalmology, University Hospital RWTH Aachen, Pauwelsstraße 30, 52074 Aachen, Germany.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.http://www.sciencedirect.com/science/article/pii/S2162253125001702MT: Delivery Strategiesnon-viral gene therapyelectroporationMüller cellsretinal degenerationrd10 mouse model |
| spellingShingle | Jiao Zhang Ziyu Bing Corinne Marie Zsuzsanna Izsvák Frank Müller Gabriele Thumann Peter Walter Sandra Johnen Effect of degeneration stage on non-viral tissue transfection of rd10 retina ex vivo Molecular Therapy: Nucleic Acids MT: Delivery Strategies non-viral gene therapy electroporation Müller cells retinal degeneration rd10 mouse model |
| title | Effect of degeneration stage on non-viral tissue transfection of rd10 retina ex vivo |
| title_full | Effect of degeneration stage on non-viral tissue transfection of rd10 retina ex vivo |
| title_fullStr | Effect of degeneration stage on non-viral tissue transfection of rd10 retina ex vivo |
| title_full_unstemmed | Effect of degeneration stage on non-viral tissue transfection of rd10 retina ex vivo |
| title_short | Effect of degeneration stage on non-viral tissue transfection of rd10 retina ex vivo |
| title_sort | effect of degeneration stage on non viral tissue transfection of rd10 retina ex vivo |
| topic | MT: Delivery Strategies non-viral gene therapy electroporation Müller cells retinal degeneration rd10 mouse model |
| url | http://www.sciencedirect.com/science/article/pii/S2162253125001702 |
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