Targeting Duchenne muscular dystrophy by skipping DMD exon 45 with base editors
Duchenne muscular dystrophy is an X-linked monogenic disease caused by mutations in the dystrophin gene (DMD) characterized by progressive muscle weakness, leading to loss of ambulation and decreased life expectancy. Since the current standard of care for Duchenne muscular dystrophy is to merely tre...
| Main Authors: | , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2023-09-01
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| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253123002020 |
| _version_ | 1797744085195292672 |
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| author | Michael Gapinske Jackson Winter Devyani Swami Lauren Gapinske Wendy S. Woods Shraddha Shirguppe Angelo Miskalis Anna Busza Dana Joulani Collin J. Kao Kurt Kostan Anne Bigot Rashid Bashir Pablo Perez-Pinera |
| author_facet | Michael Gapinske Jackson Winter Devyani Swami Lauren Gapinske Wendy S. Woods Shraddha Shirguppe Angelo Miskalis Anna Busza Dana Joulani Collin J. Kao Kurt Kostan Anne Bigot Rashid Bashir Pablo Perez-Pinera |
| author_sort | Michael Gapinske |
| collection | DOAJ |
| description | Duchenne muscular dystrophy is an X-linked monogenic disease caused by mutations in the dystrophin gene (DMD) characterized by progressive muscle weakness, leading to loss of ambulation and decreased life expectancy. Since the current standard of care for Duchenne muscular dystrophy is to merely treat symptoms, there is a dire need for treatment modalities that can correct the underlying genetic mutations. While several gene replacement therapies are being explored in clinical trials, one emerging approach that can directly correct mutations in genomic DNA is base editing. We have recently developed CRISPR-SKIP, a base editing strategy to induce permanent exon skipping by introducing C > T or A > G mutations at splice acceptors in genomic DNA, which can be used therapeutically to recover dystrophin expression when a genomic deletion leads to an out-of-frame DMD transcript. We now demonstrate that CRISPR-SKIP can be adapted to correct some forms of Duchenne muscular dystrophy by disrupting the splice acceptor in human DMD exon 45 with high efficiency, which enables open reading frame recovery and restoration of dystrophin expression. We also demonstrate that AAV-delivered split-intein base editors edit the splice acceptor of DMD exon 45 in cultured human cells and in vivo, highlighting the therapeutic potential of this strategy. |
| first_indexed | 2024-03-12T15:04:40Z |
| format | Article |
| id | doaj.art-b779d782b2df4b1795f33707a2552812 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-03-12T15:04:40Z |
| publishDate | 2023-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-b779d782b2df4b1795f33707a25528122023-08-13T04:53:36ZengElsevierMolecular Therapy: Nucleic Acids2162-25312023-09-0133572586Targeting Duchenne muscular dystrophy by skipping DMD exon 45 with base editorsMichael Gapinske0Jackson Winter1Devyani Swami2Lauren Gapinske3Wendy S. Woods4Shraddha Shirguppe5Angelo Miskalis6Anna Busza7Dana Joulani8Collin J. Kao9Kurt Kostan10Anne Bigot11Rashid Bashir12Pablo Perez-Pinera13Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USADepartment of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USADepartment of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USADepartment of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; Nick J. Holonyak Micro and Nano Technology Laboratory, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USADepartment of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USADepartment of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USADepartment of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USADepartment of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USADepartment of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USADepartment of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USADepartment of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USASorbonne Université, Inserm, Institut de Myologie, Centre de Recherche en Myologie, 75013 Paris, FranceDepartment of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; Nick J. Holonyak Micro and Nano Technology Laboratory, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; Carle Illinois College of Medicine, Champaign, IL 61820, USADepartment of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; Carle Illinois College of Medicine, Champaign, IL 61820, USA; Carl R. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; Cancer Center at Illinois, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; Department of Molecular and Integrative Physiology, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA; Corresponding author: Pablo Perez-Pinera, Department of Bioengineering, University of Illinois at Urbana-Champaign, 1406 West Green Street, Urbana 61801-2910, USA.Duchenne muscular dystrophy is an X-linked monogenic disease caused by mutations in the dystrophin gene (DMD) characterized by progressive muscle weakness, leading to loss of ambulation and decreased life expectancy. Since the current standard of care for Duchenne muscular dystrophy is to merely treat symptoms, there is a dire need for treatment modalities that can correct the underlying genetic mutations. While several gene replacement therapies are being explored in clinical trials, one emerging approach that can directly correct mutations in genomic DNA is base editing. We have recently developed CRISPR-SKIP, a base editing strategy to induce permanent exon skipping by introducing C > T or A > G mutations at splice acceptors in genomic DNA, which can be used therapeutically to recover dystrophin expression when a genomic deletion leads to an out-of-frame DMD transcript. We now demonstrate that CRISPR-SKIP can be adapted to correct some forms of Duchenne muscular dystrophy by disrupting the splice acceptor in human DMD exon 45 with high efficiency, which enables open reading frame recovery and restoration of dystrophin expression. We also demonstrate that AAV-delivered split-intein base editors edit the splice acceptor of DMD exon 45 in cultured human cells and in vivo, highlighting the therapeutic potential of this strategy.http://www.sciencedirect.com/science/article/pii/S2162253123002020MT: RNA/DNA editinggene editingCRISPR-Cas9Duchenne muscular dystrophyexon skippingadenine base editing |
| spellingShingle | Michael Gapinske Jackson Winter Devyani Swami Lauren Gapinske Wendy S. Woods Shraddha Shirguppe Angelo Miskalis Anna Busza Dana Joulani Collin J. Kao Kurt Kostan Anne Bigot Rashid Bashir Pablo Perez-Pinera Targeting Duchenne muscular dystrophy by skipping DMD exon 45 with base editors Molecular Therapy: Nucleic Acids MT: RNA/DNA editing gene editing CRISPR-Cas9 Duchenne muscular dystrophy exon skipping adenine base editing |
| title | Targeting Duchenne muscular dystrophy by skipping DMD exon 45 with base editors |
| title_full | Targeting Duchenne muscular dystrophy by skipping DMD exon 45 with base editors |
| title_fullStr | Targeting Duchenne muscular dystrophy by skipping DMD exon 45 with base editors |
| title_full_unstemmed | Targeting Duchenne muscular dystrophy by skipping DMD exon 45 with base editors |
| title_short | Targeting Duchenne muscular dystrophy by skipping DMD exon 45 with base editors |
| title_sort | targeting duchenne muscular dystrophy by skipping dmd exon 45 with base editors |
| topic | MT: RNA/DNA editing gene editing CRISPR-Cas9 Duchenne muscular dystrophy exon skipping adenine base editing |
| url | http://www.sciencedirect.com/science/article/pii/S2162253123002020 |
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