Induction of cryptic pre-mRNA splice-switching by antisense oligonucleotides

Ham, Kristin A.; Keegan, Niall; McIntosh, Craig S.; Aung-Htut, May T.; Zaw, Khine; Greer, K.; Fletcher, Sue; Wilton, Steve D.

doi:10.1038/s41598-021-94639-x

Cited by 6 publications

(4 citation statements)

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“…Eight other exons (3, 4, 5, 6, 7, 9, 10 and 11) are out-of-frame, and hence removing any of these individual exons would lead to a shift in the reading frame and may render the induced mRNA transcript susceptible to degradation through nonsense-mediated decay (NMD). However, our previous study showed that some transcripts escaped NMD when the premature stop codon was shifted to the penultimate exon after induction of targeted exon skipping 22 . Hence, in addition to the two in-frame exons, exons 2 and 8, we designed exon skipping AOs targeting exons 9, 10, or 11 to assess whether terminally truncated PCSK9 protein isoforms are produced.…”

Section: Analysis Of Pcsk9 Transcriptmentioning

confidence: 96%

Induced alternative splicing an opportunity to study PCSK9 protein isoforms at physiologically relevant concentrations

Cale,

Ham,

et al. 2023

Sci Rep

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Splice modulating antisense oligomers (AOs) are increasingly used to modulate RNA processing. While most are investigated for their use as therapeutics, AOs can also be used for basic research. This study examined their use to investigate internally and terminally truncated proprotein convertase subtilisin/kexin type 9 (PCSK9) protein isoforms. Previous studies have used plasmid or viral-vector-mediated protein overexpression to study different PCSK9 protein isoforms, creating an artificial environment within the cell. Here we designed and tested AOs to remove specific exons that encode for PCSK9 protein domains and produced protein isoforms at more physiologically relevant levels. We evaluated the isoforms’ expression, secretion, and subsequent impact on the low-density lipoprotein (LDL) receptor and its activity in Huh-7 cells. We found that modifying the Cis-His-rich domain by targeting exons 10 or 11 negatively affected LDL receptor activity and hence did not enhance LDL uptake although the levels of LDL receptor were increased. On the other hand, removing the hinge region encoded by exon 8, or a portion of the prodomain encoded by exon 2, have the potential as therapeutics for hypercholesterolemia. Our findings expand the understanding of PCSK9 isoforms and their impact on the LDL receptor and its activity at physiologically relevant concentrations.

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Section: Analysis Of Pcsk9 Transcriptmentioning

confidence: 96%

Induced alternative splicing an opportunity to study PCSK9 protein isoforms at physiologically relevant concentrations

Cale,

Ham,

et al. 2023

Sci Rep

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“…Usage of these cryptic splice sites mostly leads to a disruption of the reading frame and a reduction in the amount of functional protein produced. ssASOs can bind to the cryptic splice sites (exonic and intronic) and hide them from the splicing apparatus to restore canonical splicing [98,99]. These variants are the ideal targets for ssASOs but are less likely to be picked up with routine diagnostic sequencing and rarely have their effect reliably predicted.…”

Section: Exon Skippingmentioning

confidence: 99%

Treatability of the KMT2-Associated Neurodevelopmental Disorders Using Antisense Oligonucleotide-Based Treatments

Zardetto,

van Roon-Mom,

Aartsma-Rus

et al. 2024

Human Mutation

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Neurodevelopmental disorders (NDDs) of genetic origin are a group of early-onset neurological diseases with highly heterogeneous etiology and a symptomatic spectrum that includes intellectual disability, autism spectrum disorder, and learning and language disorders. One group of rare NDDs is associated with dysregulation of the KMT2 protein family. Members of this family share a common methyl transferase function and are involved in the etiology of rare haploinsufficiency disorders. For each of the KMT2 genes, at least one distinct disorder has been reported, yet clinical manifestations often overlap for multiple of these individually very rare disorders. Clinical care is currently focused on the management of symptoms with no targeted treatments available, illustrating a high unmet medical need and the urgency of developing disease-modifying therapeutic strategies. Antisense oligonucleotides (ASOs) are one option to treat some of these rare genetic disorders. ASOs are RNA-based treatments that can be employed to modulate gene expression through various mechanisms. In this work, we discuss the phenotypic features across the KMT2-associated NDDs and which ASO approaches are most suited for the treatment of each associated disorder. We hereby address variant-specific strategies as well as options applicable to larger groups of patients.

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“…Typically, these well-defined splice sites are recognized by the spliceosome to excise and ligate exons embedded in introns to form mature mRNA. However, mutations that interfere with authentic splicing can cause cryptic splice sites as they create a new splice site that is usually not recognized [114]. To suppress and redirect the aberrant splice site, splice-switching ASOs manipulate the splice site by binding complementarily in an antisense orientation to a specific pre-mRNA sequence and interfering with RNA duplex or normal protein-RNA interactions [115].…”

Section: Splice-switching Asosmentioning

confidence: 99%

Recent Therapeutic Progress and Future Perspectives for the Treatment of Hearing Loss

Lye,

Delaney,

Leith

et al. 2023

Biomedicines

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Up to 1.5 billion people worldwide suffer from various forms of hearing loss, with an additional 1.1 billion people at risk from various insults such as increased consumption of recreational noise-emitting devices and ageing. The most common type of hearing impairment is sensorineural hearing loss caused by the degeneration or malfunction of cochlear hair cells or spiral ganglion nerves in the inner ear. There is currently no cure for hearing loss. However, emerging frontier technologies such as gene, drug or cell-based therapies offer hope for an effective cure. In this review, we discuss the current therapeutic progress for the treatment of hearing loss. We describe and evaluate the major therapeutic approaches being applied to hearing loss and summarize the key trials and studies.

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Induction of cryptic pre-mRNA splice-switching by antisense oligonucleotides

Cited by 6 publications

References 50 publications

Induced alternative splicing an opportunity to study PCSK9 protein isoforms at physiologically relevant concentrations

Induced alternative splicing an opportunity to study PCSK9 protein isoforms at physiologically relevant concentrations

Treatability of the KMT2-Associated Neurodevelopmental Disorders Using Antisense Oligonucleotide-Based Treatments

Recent Therapeutic Progress and Future Perspectives for the Treatment of Hearing Loss

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