Comparison of CRISPR-Cas13b RNA base editing approaches for USH2A-associated inherited retinal degeneration
Abstract CRISPR-Cas13 systems have therapeutic promise for the precise correction of point mutations in RNA. Using adenosine deaminase acting on RNA (ADAR) effectors, A-I base conversions can be targeted using guide RNAs (gRNAs). We compare the Cas13 effectors PspCas13b and Cas13bt3 for the repair o...
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| Format: | Article |
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Nature Portfolio
2025-02-01
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| Series: | Communications Biology |
| Online Access: | https://doi.org/10.1038/s42003-025-07557-3 |
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| author | Lewis E. Fry Lauren Major Ahmed Salman Lucy A. McDermott Jun Yang Andrew J. King Michelle E. McClements Robert E. MacLaren |
| author_facet | Lewis E. Fry Lauren Major Ahmed Salman Lucy A. McDermott Jun Yang Andrew J. King Michelle E. McClements Robert E. MacLaren |
| author_sort | Lewis E. Fry |
| collection | DOAJ |
| description | Abstract CRISPR-Cas13 systems have therapeutic promise for the precise correction of point mutations in RNA. Using adenosine deaminase acting on RNA (ADAR) effectors, A-I base conversions can be targeted using guide RNAs (gRNAs). We compare the Cas13 effectors PspCas13b and Cas13bt3 for the repair of the gene USH2A, a common cause of inherited retinal disease and Usher syndrome. In cultured cells, we demonstrate up to 80% efficiency for the repair of the common c.11864 G > A and its murine equivalent c.11840 G > A, across different gRNAs and promoters. We develop and characterize a mouse model of Usher syndrome carrying the c.11840 G > A mutation designed for the evaluation of base editors for inherited retinal disease. Finally, we compare Cas13 effectors delivered via AAV for the repair of Ush2a in photoreceptors. Mean RNA editing rates in photoreceptors across different constructs ranged from 0.32% to 2.04%, with greater efficiency in those injected with PspCas13b compared to Cas13bt3 constructs. In mice injected with PspCas13b constructs, usherin protein was successfully restored and correctly localized to the connecting cilium following RNA editing. These results support the development of transcriptome targeting gene editing therapies for retinal disease. |
| format | Article |
| id | doaj-art-6e54696ba33c4ab7949edfbce4c0942d |
| institution | Kabale University |
| issn | 2399-3642 |
| language | English |
| publishDate | 2025-02-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Biology |
| spelling | doaj-art-6e54696ba33c4ab7949edfbce4c0942d2025-02-09T12:50:22ZengNature PortfolioCommunications Biology2399-36422025-02-018111610.1038/s42003-025-07557-3Comparison of CRISPR-Cas13b RNA base editing approaches for USH2A-associated inherited retinal degenerationLewis E. Fry0Lauren Major1Ahmed Salman2Lucy A. McDermott3Jun Yang4Andrew J. King5Michelle E. McClements6Robert E. MacLaren7Nuffield Department of Clinical Neurosciences & NIHR Oxford Biomedical Research Centre, University of OxfordNuffield Department of Clinical Neurosciences & NIHR Oxford Biomedical Research Centre, University of OxfordNuffield Department of Clinical Neurosciences & NIHR Oxford Biomedical Research Centre, University of OxfordNuffield Department of Clinical Neurosciences & NIHR Oxford Biomedical Research Centre, University of OxfordDepartment of Ophthalmology and Visual Sciences, John A. Moran Eye Center, University of UtahDepartment of Physiology, Anatomy and Genetics, University of OxfordNuffield Department of Clinical Neurosciences & NIHR Oxford Biomedical Research Centre, University of OxfordNuffield Department of Clinical Neurosciences & NIHR Oxford Biomedical Research Centre, University of OxfordAbstract CRISPR-Cas13 systems have therapeutic promise for the precise correction of point mutations in RNA. Using adenosine deaminase acting on RNA (ADAR) effectors, A-I base conversions can be targeted using guide RNAs (gRNAs). We compare the Cas13 effectors PspCas13b and Cas13bt3 for the repair of the gene USH2A, a common cause of inherited retinal disease and Usher syndrome. In cultured cells, we demonstrate up to 80% efficiency for the repair of the common c.11864 G > A and its murine equivalent c.11840 G > A, across different gRNAs and promoters. We develop and characterize a mouse model of Usher syndrome carrying the c.11840 G > A mutation designed for the evaluation of base editors for inherited retinal disease. Finally, we compare Cas13 effectors delivered via AAV for the repair of Ush2a in photoreceptors. Mean RNA editing rates in photoreceptors across different constructs ranged from 0.32% to 2.04%, with greater efficiency in those injected with PspCas13b compared to Cas13bt3 constructs. In mice injected with PspCas13b constructs, usherin protein was successfully restored and correctly localized to the connecting cilium following RNA editing. These results support the development of transcriptome targeting gene editing therapies for retinal disease.https://doi.org/10.1038/s42003-025-07557-3 |
| spellingShingle | Lewis E. Fry Lauren Major Ahmed Salman Lucy A. McDermott Jun Yang Andrew J. King Michelle E. McClements Robert E. MacLaren Comparison of CRISPR-Cas13b RNA base editing approaches for USH2A-associated inherited retinal degeneration Communications Biology |
| title | Comparison of CRISPR-Cas13b RNA base editing approaches for USH2A-associated inherited retinal degeneration |
| title_full | Comparison of CRISPR-Cas13b RNA base editing approaches for USH2A-associated inherited retinal degeneration |
| title_fullStr | Comparison of CRISPR-Cas13b RNA base editing approaches for USH2A-associated inherited retinal degeneration |
| title_full_unstemmed | Comparison of CRISPR-Cas13b RNA base editing approaches for USH2A-associated inherited retinal degeneration |
| title_short | Comparison of CRISPR-Cas13b RNA base editing approaches for USH2A-associated inherited retinal degeneration |
| title_sort | comparison of crispr cas13b rna base editing approaches for ush2a associated inherited retinal degeneration |
| url | https://doi.org/10.1038/s42003-025-07557-3 |
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