Correction of the Splicing Defect Caused by a Recurrent Variant in <i>ABCA4</i> (c.769-784C>T) That Underlies Stargardt Disease
Stargardt disease is an inherited retinal disease caused by biallelic mutations in the <i>ABCA4</i> gene, many of which affect <i>ABCA4</i> splicing. In this study, nine antisense oligonucleotides (AONs) were designed to correct pseudoexon (PE) inclusion caused by a recurrent...
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MDPI AG
2022-12-01
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| Online Access: | https://www.mdpi.com/2073-4409/11/24/3947 |
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| author | Tomasz Z. Tomkiewicz Sara E. Nieuwenhuis Frans P. M. Cremers Alejandro Garanto Rob W. J. Collin |
| author_facet | Tomasz Z. Tomkiewicz Sara E. Nieuwenhuis Frans P. M. Cremers Alejandro Garanto Rob W. J. Collin |
| author_sort | Tomasz Z. Tomkiewicz |
| collection | DOAJ |
| description | Stargardt disease is an inherited retinal disease caused by biallelic mutations in the <i>ABCA4</i> gene, many of which affect <i>ABCA4</i> splicing. In this study, nine antisense oligonucleotides (AONs) were designed to correct pseudoexon (PE) inclusion caused by a recurrent deep-intronic variant in <i>ABCA4</i> (c.769-784C>T). First, the ability of AONs to skip the PE from the final <i>ABCA4</i> mRNA transcript was assessed in two cellular models carrying the c.769-784C>T variant: a midigene assay using HEK293T cells and patient-derived fibroblasts. Based on the splicing-correcting ability of each individual AON, the three most efficacious AONs targeting independent regions of the PE were selected for a final assessment in photoreceptor precursor cells (PPCs). The final analysis in the PPC model confirmed high efficacy of AON2, -5, and -7 in promoting PE exclusion. Among the three AONs, AON2 is chosen as the lead candidate for further optimization, hereby showcasing the high potential of AONs to correct aberrant splicing events driven by deep-intronic variants. |
| first_indexed | 2024-03-09T17:13:45Z |
| format | Article |
| id | doaj.art-d332dd9035664406962ab8615069c299 |
| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-09T17:13:45Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj.art-d332dd9035664406962ab8615069c2992023-11-24T13:53:11ZengMDPI AGCells2073-44092022-12-011124394710.3390/cells11243947Correction of the Splicing Defect Caused by a Recurrent Variant in <i>ABCA4</i> (c.769-784C>T) That Underlies Stargardt DiseaseTomasz Z. Tomkiewicz0Sara E. Nieuwenhuis1Frans P. M. Cremers2Alejandro Garanto3Rob W. J. Collin4Department of Human Genetics and Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, 6525 GA Nijmegen, The NetherlandsDepartment of Human Genetics and Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, 6525 GA Nijmegen, The NetherlandsDepartment of Human Genetics and Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, 6525 GA Nijmegen, The NetherlandsDepartments of Pediatrics, Amalia Children’s Hospital, Human Genetics and Radboud Institute for Molecular Life Sciences, Radboud University Medical Center, 6525 GA Nijmegen, The NetherlandsDepartment of Human Genetics and Donders Institute for Brain, Cognition and Behaviour, Radboud University Medical Center, 6525 GA Nijmegen, The NetherlandsStargardt disease is an inherited retinal disease caused by biallelic mutations in the <i>ABCA4</i> gene, many of which affect <i>ABCA4</i> splicing. In this study, nine antisense oligonucleotides (AONs) were designed to correct pseudoexon (PE) inclusion caused by a recurrent deep-intronic variant in <i>ABCA4</i> (c.769-784C>T). First, the ability of AONs to skip the PE from the final <i>ABCA4</i> mRNA transcript was assessed in two cellular models carrying the c.769-784C>T variant: a midigene assay using HEK293T cells and patient-derived fibroblasts. Based on the splicing-correcting ability of each individual AON, the three most efficacious AONs targeting independent regions of the PE were selected for a final assessment in photoreceptor precursor cells (PPCs). The final analysis in the PPC model confirmed high efficacy of AON2, -5, and -7 in promoting PE exclusion. Among the three AONs, AON2 is chosen as the lead candidate for further optimization, hereby showcasing the high potential of AONs to correct aberrant splicing events driven by deep-intronic variants.https://www.mdpi.com/2073-4409/11/24/3947antisense oligonucleotides<i>ABCA4</i>Stargardt diseaseinherited retinal diseasesplicing modulationRNA therapy |
| spellingShingle | Tomasz Z. Tomkiewicz Sara E. Nieuwenhuis Frans P. M. Cremers Alejandro Garanto Rob W. J. Collin Correction of the Splicing Defect Caused by a Recurrent Variant in <i>ABCA4</i> (c.769-784C>T) That Underlies Stargardt Disease Cells antisense oligonucleotides <i>ABCA4</i> Stargardt disease inherited retinal disease splicing modulation RNA therapy |
| title | Correction of the Splicing Defect Caused by a Recurrent Variant in <i>ABCA4</i> (c.769-784C>T) That Underlies Stargardt Disease |
| title_full | Correction of the Splicing Defect Caused by a Recurrent Variant in <i>ABCA4</i> (c.769-784C>T) That Underlies Stargardt Disease |
| title_fullStr | Correction of the Splicing Defect Caused by a Recurrent Variant in <i>ABCA4</i> (c.769-784C>T) That Underlies Stargardt Disease |
| title_full_unstemmed | Correction of the Splicing Defect Caused by a Recurrent Variant in <i>ABCA4</i> (c.769-784C>T) That Underlies Stargardt Disease |
| title_short | Correction of the Splicing Defect Caused by a Recurrent Variant in <i>ABCA4</i> (c.769-784C>T) That Underlies Stargardt Disease |
| title_sort | correction of the splicing defect caused by a recurrent variant in i abca4 i c 769 784c t that underlies stargardt disease |
| topic | antisense oligonucleotides <i>ABCA4</i> Stargardt disease inherited retinal disease splicing modulation RNA therapy |
| url | https://www.mdpi.com/2073-4409/11/24/3947 |
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