AI Article Synopsis
- Antisense oligonucleotides (AONs) can affect mRNA processing by causing degradation, stopping translation, or changing splicing patterns, depending on their design and properties.
- A study of 156 AONs aimed at modifying the dystrophin transcript found that successful AONs had higher guanine-cytosine content and stronger binding energies, and were closer to splice sites compared to ineffective ones.
- The research identified key factors that contribute to AON effectiveness, enabling a classification accuracy of 79%, and offers guidelines for future AON design to enhance splice modulation.
Article Abstract
Antisense oligonucleotides (AONs) can interfere with mRNA processing through RNase H-mediated degradation, translational arrest, or modulation of splicing. The antisense approach relies on AONs to efficiently bind to target sequences and depends on AON length, sequence content, secondary structure, thermodynamic properties, and target accessibility. We here performed a retrospective analysis of a series of 156 AONs (104 effective, 52 ineffective) previously designed and evaluated for splice modulation of the dystrophin transcript. This showed that the guanine-cytosine content and the binding energies of AON-target and AON-AON complexes were significantly higher for effective AONs. Effective AONs were also located significantly closer to the acceptor splice site (SS). All analyzed AONs are exon-internal and may act through steric hindrance of Ser-Arg-rich (SR) proteins to exonic splicing enhancer (ESE) sites. Indeed, effective AONs were significantly enriched for ESEs predicted by ESE software programs, except for predicted binding sites of SR protein Tra2beta, which were significantly enriched in ineffective AONs. These findings compile guidelines for development of AONs and provide more insight into the mechanism of antisense-mediated exon skipping. On the basis of only four parameters, we could correctly classify 79% of all AONs as effective or ineffective, suggesting these parameters can be used to more optimally design splice-modulating AONs.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2835096 | PMC |
| http://dx.doi.org/10.1038/mt.2008.205 | DOI Listing |
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