Single Exon Skipping Can Address a Multi-Exon Duplication in the Dystrophin Gene
| Autor: | Yoram Nevo, Russell D. Johnsen, Steve D. Wilton, K. Greer, Yakov Fellig, Sue Fletcher |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
musculoskeletal diseases
Duchenne muscular dystrophy congenital hereditary and neonatal diseases and abnormalities RNA Splicing Oligonucleotides Catalysis Article dystrophin Inorganic Chemistry lcsh:Chemistry Exon splicing INDEL Mutation duplication mutations Gene duplication medicine Humans antisense oligomers Physical and Theoretical Chemistry Muscle Skeletal Molecular Biology lcsh:QH301-705.5 Spectroscopy Cells Cultured Genetics Messenger RNA biology Organic Chemistry General Medicine Exons Genetic Therapy Oligonucleotides Antisense medicine.disease Exon skipping Computer Science Applications Blot Muscular Dystrophy Duchenne lcsh:Biology (General) lcsh:QD1-999 RNA splicing biology.protein Dystrophin exon skipping |
| Zdroj: | International Journal of Molecular Sciences International Journal of Molecular Sciences, Vol 21, Iss 4511, p 4511 (2020) Volume 21 Issue 12 |
| ISSN: | 1422-0067 |
| Popis: | Duchenne muscular dystrophy (DMD) is a severe muscle wasting disease typically caused by protein-truncating mutations that preclude synthesis of a functional dystrophin. Exonic deletions are the most common type of DMD lesion, however, whole exon duplications account for between 10&ndash 15% of all reported mutations. Here, we describe in vitro evaluation of antisense oligonucleotide-induced splice switching strategies to re-frame the transcript disrupted by a multi-exon duplication within the DMD gene. Phosphorodiamidate morpholino oligomers and phosphorodiamidate morpholino oligomers coupled to a cell penetrating peptide were evaluated in a Duchenne muscular dystrophy patient cell strain carrying an exon 14&ndash 17 duplication. Two strategies were employed the conventional approach was to remove both copies of exon 17 in addition to exon 18, and the second strategy was to remove only the first copy of exon 17. Both approaches result in a larger than normal but in-frame DMD transcript, but surprisingly, the removal of only the first exon 17 appeared to be more efficient in restoring dystrophin, as determined using western blotting. The emergence of a normal sized DMD mRNA transcript that was not apparent in untreated samples may have arisen from back splicing and could also account for some of the dystrophin protein being produced. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|