Unknown

Dataset Information

0

Successful large gene augmentation of USH2A with non-viral episomal vectors.


ABSTRACT: USH2A mutations are a common cause of autosomal recessive retinitis pigmentosa (RP) and Usher syndrome, for which there are currently no approved treatments. Gene augmentation is a valuable therapeutic strategy for treating many inherited retinal diseases; however, conventional adeno-associated virus (AAV) gene therapy cannot accommodate cDNAs exceeding 4.7 kb, such as the 15.6-kb-long USH2A coding sequence. In the present study, we adopted an alternative strategy to successfully generate scaffold/matrix attachment region (S/MAR) DNA plasmid vectors containing the full-length human USH2A coding sequence, a GFP reporter gene, and a ubiquitous promoter (CMV or CAG), reaching a size of approximately 23 kb. We assessed the vectors in transfected HEK293 cells and USH2A patient-derived dermal fibroblasts in addition to ush2au507 zebrafish microinjected with the vector at the one-cell stage. pS/MAR-USH2A vectors drove persistent transgene expression in patient fibroblasts with restoration of usherin. Twelve months of GFP expression was detected in the photoreceptor cells, with rescue of Usher 2 complex localization in the photoreceptors of ush2au507 zebrafish retinas injected with pS/MAR-USH2A. To our knowledge, this is the first reported vector that can be used to express full-length usherin with functional rescue. S/MAR DNA vectors have shown promise as a novel non-viral retinal gene therapy, warranting further translational development.

SUBMITTER: Toms M 

PROVIDER: S-EPMC10491995 | biostudies-literature | 2023 Sep

REPOSITORIES: biostudies-literature

altmetric image

Publications

Successful large gene augmentation of USH2A with non-viral episomal vectors.

Toms Maria M   Toualbi Lyes L   Almeida Patrick V PV   Harbottle Richard R   Moosajee Mariya M  

Molecular therapy : the journal of the American Society of Gene Therapy 20230619 9


USH2A mutations are a common cause of autosomal recessive retinitis pigmentosa (RP) and Usher syndrome, for which there are currently no approved treatments. Gene augmentation is a valuable therapeutic strategy for treating many inherited retinal diseases; however, conventional adeno-associated virus (AAV) gene therapy cannot accommodate cDNAs exceeding 4.7 kb, such as the 15.6-kb-long USH2A coding sequence. In the present study, we adopted an alternative strategy to successfully generate scaffo  ...[more]

Similar Datasets

| S-EPMC6359729 | biostudies-literature
| S-EPMC4195477 | biostudies-literature
| S-EPMC3413895 | biostudies-literature
| S-EPMC5019568 | biostudies-literature
| S-EPMC7956638 | biostudies-literature
| S-EPMC6334614 | biostudies-literature
| S-EPMC9502120 | biostudies-literature
| S-EPMC10530965 | biostudies-literature
| S-EPMC6023384 | biostudies-literature
| S-EPMC8743282 | biostudies-literature