Repair of rhodopsin mRNA by spliceosome-mediated RNA trans-splicing: a new approach for autosomal dominant retinitis pigmentosa.

Details

Serval ID
serval:BIB_23C134DDA937
Type
Article: article from journal or magazin.
Collection
Publications
Title
Repair of rhodopsin mRNA by spliceosome-mediated RNA trans-splicing: a new approach for autosomal dominant retinitis pigmentosa.
Journal
Molecular therapy
Author(s)
Berger A., Lorain S., Joséphine C., Desrosiers M., Peccate C., Voit T., Garcia L., Sahel J.A., Bemelmans A.P.
ISSN
1525-0024 (Electronic)
ISSN-L
1525-0016
Publication state
Published
Issued date
05/2015
Peer-reviewed
Oui
Volume
23
Number
5
Pages
918-930
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
The promising clinical results obtained for ocular gene therapy in recent years have paved the way for gene supplementation to treat recessively inherited forms of retinal degeneration. The situation is more complex for dominant mutations, as the toxic mutant gene product must be removed. We used spliceosome-mediated RNA trans-splicing as a strategy for repairing the transcript of the rhodopsin gene, the gene most frequently mutated in autosomal dominant retinitis pigmentosa. We tested 17 different molecules targeting the pre-mRNA intron 1, by transient transfection of HEK-293T cells, with subsequent trans-splicing quantification at the transcript level. We found that the targeting of some parts of the intron promoted trans-splicing more efficiently than the targeting of other areas, and that trans-splicing rate could be increased by modifying the replacement sequence. We then developed cell lines stably expressing the rhodopsin gene, for the assessment of phenotypic criteria relevant to the pathogenesis of retinitis pigmentosa. Using this model, we showed that trans-splicing restored the correct localization of the protein to the plasma membrane. Finally, we tested our best candidate by AAV gene transfer in a mouse model of retinitis pigmentosa that expresses a mutant allele of the human rhodopsin gene, and demonstrated the feasibility of trans-splicing in vivo. This work paves the way for trans-splicing gene therapy to treat retinitis pigmentosa due to rhodopsin gene mutation and, more generally, for the treatment of genetic diseases with dominant transmission.
Keywords
Animals, Binding Sites, Cell Line, Transformed, Gene Expression, Genes, Dominant, Genetic Therapy, Genetic Vectors/genetics, Humans, Introns, Mice, Mice, Transgenic, Phenotype, Photoreceptor Cells/metabolism, Protein Interaction Domains and Motifs/genetics, Protein Transport, RNA Precursors/genetics, RNA Splicing, RNA, Messenger/genetics, Retinitis Pigmentosa/genetics, Retinitis Pigmentosa/therapy, Rhodopsin/chemistry, Rhodopsin/genetics, Rhodopsin/metabolism, Trans-Splicing, Transduction, Genetic
Pubmed
Web of science
Open Access
Yes
Create date
15/07/2020 10:08
Last modification date
16/07/2020 8:43
Usage data