Visual function restoration in a mouse model of Leber congenital amaurosis via therapeutic base editing
Leber congenital amaurosis (LCA), an inherited retinal degeneration, causes severe visual dysfunction in children and adolescents. In patients with LCA, pathogenic variants, such as RPE65, are evident in specific genes, related to the functions of retinal pigment epithelium and photoreceptors. In co...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2023-03-01
|
| Series: | Molecular Therapy: Nucleic Acids |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2162253122003134 |
| _version_ | 1828116842015621120 |
|---|---|
| author | Dong Hyun Jo Hyeon-Ki Jang Chang Sik Cho Jun Hee Han Gahee Ryu Youngri Jung Sangsu Bae Jeong Hun Kim |
| author_facet | Dong Hyun Jo Hyeon-Ki Jang Chang Sik Cho Jun Hee Han Gahee Ryu Youngri Jung Sangsu Bae Jeong Hun Kim |
| author_sort | Dong Hyun Jo |
| collection | DOAJ |
| description | Leber congenital amaurosis (LCA), an inherited retinal degeneration, causes severe visual dysfunction in children and adolescents. In patients with LCA, pathogenic variants, such as RPE65, are evident in specific genes, related to the functions of retinal pigment epithelium and photoreceptors. In contrast to the original Cas9, base editing tools can correct pathogenic substitutions without generation of DNA double-stranded breaks (DSBs). In this study, dual adeno-associated virus (AAV) vectors containing split adenine base editors (ABEs) with trans-splicing intein were prepared for in vivo base editing in retinal degeneration of 12 (rd12) mice, an animal model of LCA, possessing a nonsense mutation of C to T transition in the Rpe65 gene (p.R44X). Subretinal injection of AAV-ABE in retinal pigment epithelial cells of rd12 mice resulted in an A to G transition. The on-target editing was sufficient for recovery of wild-type mRNA, RPE65 protein, and light-induced electrical responses from the retina. Compared with our previous therapeutic editing strategies using Cas9 and prime editing, or with the gene transfer strategy shown in the current study, our results suggest that, considering the editing efficacy and functional recovery, ABEs could be a strong, reliable method for correction of pathogenic variants in the treatment of LCA. |
| first_indexed | 2024-04-11T13:04:33Z |
| format | Article |
| id | doaj.art-0e40f671c79f40dd9e740725b178edd1 |
| institution | Directory Open Access Journal |
| issn | 2162-2531 |
| language | English |
| last_indexed | 2024-04-11T13:04:33Z |
| publishDate | 2023-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Nucleic Acids |
| spelling | doaj.art-0e40f671c79f40dd9e740725b178edd12022-12-22T04:22:50ZengElsevierMolecular Therapy: Nucleic Acids2162-25312023-03-01311627Visual function restoration in a mouse model of Leber congenital amaurosis via therapeutic base editingDong Hyun Jo0Hyeon-Ki Jang1Chang Sik Cho2Jun Hee Han3Gahee Ryu4Youngri Jung5Sangsu Bae6Jeong Hun Kim7Department of Anatomy and Cell Biology, Seoul National University College of Medicine, Seoul 03080, Republic of KoreaDivision of Chemical Engineering and Bioengineering, College of Art Culture and Engineering, Kangwon National University, Chuncheon 24341, Republic of KoreaFight Against Angiogenesis-Related Blindness (FARB) Laboratory, Biomedical Research Institute, Seoul National University Hospital, Seoul 03080, Republic of KoreaDepartment of Chemistry, Hanyang University, Seoul, Republic of KoreaDepartment of Chemistry, Hanyang University, Seoul, Republic of KoreaDepartment of Chemistry, Hanyang University, Seoul, Republic of KoreaDepartment of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Republic of Korea; Cancer Research Institute, Seoul National University, Seoul 03080, Republic of Korea; Corresponding author Sangsu Bae, PhD, Department of Biomedical Sciences, and Department of Biochemistry and Molecular Biology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.Fight Against Angiogenesis-Related Blindness (FARB) Laboratory, Biomedical Research Institute, Seoul National University Hospital, Seoul 03080, Republic of Korea; Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Republic of Korea; Department of Ophthalmology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea; Advanced Biomedical Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB), Daejeon 34141, Republic of Korea; Institute of Reproductive Medicine and Population, Seoul National University College of Medicine, Seoul 03080, Republic of Korea; Corresponding author Jeong Hun Kim, MD, PhD, Fight against Angiogenesis-Related Blindness (FARB) Laboratory, Biomedical Research Institute, Seoul National University Hospital, and Department of Ophthalmology, Seoul National University College of Medicine, Seoul 03080, Republic of Korea.Leber congenital amaurosis (LCA), an inherited retinal degeneration, causes severe visual dysfunction in children and adolescents. In patients with LCA, pathogenic variants, such as RPE65, are evident in specific genes, related to the functions of retinal pigment epithelium and photoreceptors. In contrast to the original Cas9, base editing tools can correct pathogenic substitutions without generation of DNA double-stranded breaks (DSBs). In this study, dual adeno-associated virus (AAV) vectors containing split adenine base editors (ABEs) with trans-splicing intein were prepared for in vivo base editing in retinal degeneration of 12 (rd12) mice, an animal model of LCA, possessing a nonsense mutation of C to T transition in the Rpe65 gene (p.R44X). Subretinal injection of AAV-ABE in retinal pigment epithelial cells of rd12 mice resulted in an A to G transition. The on-target editing was sufficient for recovery of wild-type mRNA, RPE65 protein, and light-induced electrical responses from the retina. Compared with our previous therapeutic editing strategies using Cas9 and prime editing, or with the gene transfer strategy shown in the current study, our results suggest that, considering the editing efficacy and functional recovery, ABEs could be a strong, reliable method for correction of pathogenic variants in the treatment of LCA.http://www.sciencedirect.com/science/article/pii/S2162253122003134MT: RNA/DNA editingbase editingCRISPRLeber congenital amaurosisRPE65adeno-associated virus |
| spellingShingle | Dong Hyun Jo Hyeon-Ki Jang Chang Sik Cho Jun Hee Han Gahee Ryu Youngri Jung Sangsu Bae Jeong Hun Kim Visual function restoration in a mouse model of Leber congenital amaurosis via therapeutic base editing Molecular Therapy: Nucleic Acids MT: RNA/DNA editing base editing CRISPR Leber congenital amaurosis RPE65 adeno-associated virus |
| title | Visual function restoration in a mouse model of Leber congenital amaurosis via therapeutic base editing |
| title_full | Visual function restoration in a mouse model of Leber congenital amaurosis via therapeutic base editing |
| title_fullStr | Visual function restoration in a mouse model of Leber congenital amaurosis via therapeutic base editing |
| title_full_unstemmed | Visual function restoration in a mouse model of Leber congenital amaurosis via therapeutic base editing |
| title_short | Visual function restoration in a mouse model of Leber congenital amaurosis via therapeutic base editing |
| title_sort | visual function restoration in a mouse model of leber congenital amaurosis via therapeutic base editing |
| topic | MT: RNA/DNA editing base editing CRISPR Leber congenital amaurosis RPE65 adeno-associated virus |
| url | http://www.sciencedirect.com/science/article/pii/S2162253122003134 |
| work_keys_str_mv | AT donghyunjo visualfunctionrestorationinamousemodeloflebercongenitalamaurosisviatherapeuticbaseediting AT hyeonkijang visualfunctionrestorationinamousemodeloflebercongenitalamaurosisviatherapeuticbaseediting AT changsikcho visualfunctionrestorationinamousemodeloflebercongenitalamaurosisviatherapeuticbaseediting AT junheehan visualfunctionrestorationinamousemodeloflebercongenitalamaurosisviatherapeuticbaseediting AT gaheeryu visualfunctionrestorationinamousemodeloflebercongenitalamaurosisviatherapeuticbaseediting AT youngrijung visualfunctionrestorationinamousemodeloflebercongenitalamaurosisviatherapeuticbaseediting AT sangsubae visualfunctionrestorationinamousemodeloflebercongenitalamaurosisviatherapeuticbaseediting AT jeonghunkim visualfunctionrestorationinamousemodeloflebercongenitalamaurosisviatherapeuticbaseediting |