Dystrophin rescue by trans-splicing: a strategy for DMD genotypes not eligible for exon skipping approaches
| Autor: | Graziella Griffith, Guillaume Precigout, Stéphanie Lorain, Luis Garcia, Thomas Voit, Maëva Le Hir, Susanne Philippi, Arnaud Jollet, Cécile Peccate, Kamel Mamchaoui |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
musculoskeletal diseases
mdx mouse congenital hereditary and neonatal diseases and abnormalities Genotype Duchenne muscular dystrophy Genetic Vectors Muscle Fibers Skeletal Trans-Splicing Dystrophin Exon Mice Genetics medicine Animals Humans RNA Messenger biology Muscles MRNA stabilization Exons Dependovirus medicine.disease Exon skipping Introns Muscular Dystrophy Duchenne RNA splicing Synthetic Biology and Chemistry biology.protein Mice Inbred mdx NIH 3T3 Cells RNA Splice Sites Minigene |
| Zdroj: | Nucleic Acids Research |
| ISSN: | 1362-4962 0305-1048 |
| Popis: | RNA-based therapeutic approaches using splice-switching oligonucleotides have been successfully applied to rescue dystrophin in Duchenne muscular dystrophy (DMD) preclinical models and are currently being evaluated in DMD patients. Although the modular structure of dystrophin protein tolerates internal deletions, many mutations that affect nondispensable domains of the protein require further strategies. Among these, trans-splicing technology is particularly attractive, as it allows the replacement of any mutated exon by its normal version as well as introducing missing exons or correcting duplication mutations. We have applied such a strategy in vitro by using cotransfection of pre-trans-splicing molecule (PTM) constructs along with a reporter minigene containing part of the dystrophin gene harboring the stop-codon mutation found in the mdx mouse model of DMD. Optimization of the different functional domains of the PTMs allowed achieving accurate and efficient trans-splicing of up to 30% of the transcript encoded by the cotransfected minigene. Optimized parameters included mRNA stabilization, choice of splice site sequence, inclusion of exon splice enhancers and artificial intronic sequence. Intramuscular delivery of adeno-associated virus vectors expressing PTMs allowed detectable levels of dystrophin in mdx and mdx4Cv, illustrating that a given PTM can be suitable for a variety of mutations. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|