Rhodopsin Genomic Loci DNA Nanoparticles Improve Expression and Rescue of Retinal Degeneration in a Model for Retinitis Pigmentosa

The use of gene therapy may allow replacement of the defective gene. Minigenes, such as cDNAs, are often used. However, these may not express normal physiological genetic profiles due to lack of crucial endogenous regulatory elements. We constructed DNA nanoparticles (NPs) that contain either the mo...

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Bibliographic Details
Published in:Molecular therapy 2020-02, Vol.28 (2), p.523-535
Main Authors: Zheng, Min, Mitra, Rajendra N., Weiss, Ellen R., Han, Zongchao
Format: Article
Language:English
Subjects:
Animals
Disease Models, Animal
DNA - administration & dosage
Endoplasmic Reticulum Stress
full-length genomic DNA
gene delivery
Gene Expression
Gene Transfer Techniques
Genetic Therapy
Humans
Mice
Mice, Knockout
Nanoparticles
Original
Photoreceptor Cells, Vertebrate - metabolism
physiological transgene expression
Retinal Degeneration - genetics
Retinal Degeneration - pathology
Retinal Degeneration - therapy
retinitis pigmentosa
Retinitis Pigmentosa - genetics
Retinitis Pigmentosa - pathology
Retinitis Pigmentosa - therapy
Rhodopsin - genetics
Transgenes
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Summary:The use of gene therapy may allow replacement of the defective gene. Minigenes, such as cDNAs, are often used. However, these may not express normal physiological genetic profiles due to lack of crucial endogenous regulatory elements. We constructed DNA nanoparticles (NPs) that contain either the mouse or human full-length rhodopsin genomic locus, including endogenous promoters, all introns, and flanking regulatory sequences of the 15–16 kb genomic rhodopsin DNA inserts. We transduced the NPs into primary retinal cell cultures from the rhodopsin knockout (RKO) mouse in vitro and into the RKO mouse in vivo and compared the effects on different functions to plasmid cDNA NP counterparts that were driven by ubiquitous promoters. Our results demonstrate that genomic DNA vectors resulted in long-term high levels of physiological transgene expression over a period of 5 months. In contrast, the cDNA counterparts exhibited low levels of expression with sensitivity to the endoplasmic reticulum (ER) stress mechanism using the same transgene copy number both in vitro and in vivo. This study demonstrates for the first time the transducing of the rhodopsin genomic locus using compacted DNA NPs. Zheng et al. demonstrated that delivery of DNA nanoparticles containing the entire genomic transgene locus of rhodopsin can potentially pass the endoplasmic reticulum quality control and rescue retinal deficits in rhodopsin knockout mouse eyes. Given that all endogenous elements were included in the transgene, their work has a positive impact on physiological transgene expression for improving gene therapy.
ISSN:1525-0016
1525-0024
DOI:10.1016/j.ymthe.2019.11.031