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Antisense targeting of decoy exons can reduce intron retention and increase protein expression in human erythroblasts
The decoy exon model has been proposed to regulate a subset of intron retention (IR) events involving predominantly larger introns (>1 kb). Splicing reporter studies have shown that decoy splice sites are essential for activity, suggesting that decoys act by engaging intron-terminal splice sites...
Autores principales: | , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory Press
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7373989/ https://www.ncbi.nlm.nih.gov/pubmed/32312846 http://dx.doi.org/10.1261/rna.075028.120 |
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author | Parra, Marilyn Zhang, Weiguo Vu, Jonathan DeWitt, Mark Conboy, John G. |
author_facet | Parra, Marilyn Zhang, Weiguo Vu, Jonathan DeWitt, Mark Conboy, John G. |
author_sort | Parra, Marilyn |
collection | PubMed |
description | The decoy exon model has been proposed to regulate a subset of intron retention (IR) events involving predominantly larger introns (>1 kb). Splicing reporter studies have shown that decoy splice sites are essential for activity, suggesting that decoys act by engaging intron-terminal splice sites and competing with cross-intron interactions required for intron excision. The decoy model predicts that antisense oligonucleotides may be able to block decoy splice sites in endogenous pre-mRNA, thereby reducing IR and increasing productive gene expression. Indeed, we now demonstrate that targeting a decoy 5′ splice site in the O-GlcNAc transferase (OGT) gene reduced IR from ∼80% to ∼20% in primary human erythroblasts, accompanied by increases in spliced OGT RNA and OGT protein expression. The remaining OGT IR was refractory to antisense treatment and might be mediated by independent mechanism(s). In contrast, other retained introns were strongly dependent on decoy function, since antisense targeting of decoy 5′ splice sites greatly reduced (SNRNP70) or nearly eliminated (SF3B1) IR in two widely expressed splicing factors, and also greatly reduced IR in transcripts encoding the erythroid-specific structural protein, α-spectrin (SPTA1). These results show that modulating decoy exon function can dramatically alter IR and suggest that dynamic regulation of decoy exons could be a mechanism to fine-tune gene expression post-transcriptionally in many cell types. |
format | Online Article Text |
id | pubmed-7373989 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-73739892021-08-01 Antisense targeting of decoy exons can reduce intron retention and increase protein expression in human erythroblasts Parra, Marilyn Zhang, Weiguo Vu, Jonathan DeWitt, Mark Conboy, John G. RNA Article The decoy exon model has been proposed to regulate a subset of intron retention (IR) events involving predominantly larger introns (>1 kb). Splicing reporter studies have shown that decoy splice sites are essential for activity, suggesting that decoys act by engaging intron-terminal splice sites and competing with cross-intron interactions required for intron excision. The decoy model predicts that antisense oligonucleotides may be able to block decoy splice sites in endogenous pre-mRNA, thereby reducing IR and increasing productive gene expression. Indeed, we now demonstrate that targeting a decoy 5′ splice site in the O-GlcNAc transferase (OGT) gene reduced IR from ∼80% to ∼20% in primary human erythroblasts, accompanied by increases in spliced OGT RNA and OGT protein expression. The remaining OGT IR was refractory to antisense treatment and might be mediated by independent mechanism(s). In contrast, other retained introns were strongly dependent on decoy function, since antisense targeting of decoy 5′ splice sites greatly reduced (SNRNP70) or nearly eliminated (SF3B1) IR in two widely expressed splicing factors, and also greatly reduced IR in transcripts encoding the erythroid-specific structural protein, α-spectrin (SPTA1). These results show that modulating decoy exon function can dramatically alter IR and suggest that dynamic regulation of decoy exons could be a mechanism to fine-tune gene expression post-transcriptionally in many cell types. Cold Spring Harbor Laboratory Press 2020-08 /pmc/articles/PMC7373989/ /pubmed/32312846 http://dx.doi.org/10.1261/rna.075028.120 Text en © 2020 Parra et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/. |
spellingShingle | Article Parra, Marilyn Zhang, Weiguo Vu, Jonathan DeWitt, Mark Conboy, John G. Antisense targeting of decoy exons can reduce intron retention and increase protein expression in human erythroblasts |
title | Antisense targeting of decoy exons can reduce intron retention and increase protein expression in human erythroblasts |
title_full | Antisense targeting of decoy exons can reduce intron retention and increase protein expression in human erythroblasts |
title_fullStr | Antisense targeting of decoy exons can reduce intron retention and increase protein expression in human erythroblasts |
title_full_unstemmed | Antisense targeting of decoy exons can reduce intron retention and increase protein expression in human erythroblasts |
title_short | Antisense targeting of decoy exons can reduce intron retention and increase protein expression in human erythroblasts |
title_sort | antisense targeting of decoy exons can reduce intron retention and increase protein expression in human erythroblasts |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7373989/ https://www.ncbi.nlm.nih.gov/pubmed/32312846 http://dx.doi.org/10.1261/rna.075028.120 |
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