Journal article
Systematic approach to developing splice modulating antisense oligonucleotides
International Journal of Molecular Sciences, Vol.20(20), Article 5030
2019
Abstract
The process of pre-mRNA splicing is a common and fundamental step in the expression of most human genes. Alternative splicing, whereby different splice motifs and sites are recognised in a developmental and/or tissue-specific manner, contributes to genetic plasticity and diversity of gene expression. Redirecting pre-mRNA processing of various genes has now been validated as a viable clinical therapeutic strategy, providing treatments for Duchenne muscular dystrophy (inducing specific exon skipping) and spinal muscular atrophy (promoting exon retention). We have designed and evaluated over 5000 different antisense oligonucleotides to alter splicing of a variety of pre-mRNAs, from the longest known human pre-mRNA to shorter, exon-dense primary gene transcripts. Here, we present our guidelines for designing, evaluating and optimising splice switching antisense oligomers in vitro. These systematic approaches assess several critical factors such as the selection of target splicing motifs, choice of cells, various delivery reagents and crucial aspects of validating assays for the screening of antisense oligonucleotides composed of 2′-O-methyl modified bases on a phosphorothioate backbone.
Details
- Title
- Systematic approach to developing splice modulating antisense oligonucleotides
- Authors/Creators
- M.T. Aung-Htut (Author/Creator) - Murdoch UniversityC.S. McIntosh (Author/Creator) - Murdoch UniversityK.A. Ham (Author/Creator) - Murdoch UniversityI.L. Pitout (Author/Creator) - Murdoch UniversityL.L. Flynn (Author/Creator) - Murdoch UniversityK. Greer (Author/Creator) - Murdoch UniversityS. Fletcher (Author/Creator) - Murdoch UniversityS.D. Wilton (Author/Creator) - Murdoch University
- Publication Details
- International Journal of Molecular Sciences, Vol.20(20), Article 5030
- Publisher
- MDPI
- Identifiers
- 991005540137507891
- Copyright
- © 2019 by the authors. Licensee MDPI, Basel, Switzerland.
- Murdoch Affiliation
- Centre for Molecular Medicine and Innovative Therapeutics
- Language
- English
- Resource Type
- Journal article
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- Collaboration types
- Domestic collaboration
- Citation topics
- 1 Clinical & Life Sciences
- 1.255 Musculoskeletal Disorders
- 1.255.628 Duchenne Muscular Dystrophy
- Web Of Science research areas
- Biochemistry & Molecular Biology
- Chemistry, Multidisciplinary
- ESI research areas
- Chemistry