{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T00:50:16Z","timestamp":1782867016815,"version":"3.54.5"},"reference-count":71,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2024,5,15]],"date-time":"2024-05-15T00:00:00Z","timestamp":1715731200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"UKRI MRC"},{"name":"NIHR Oxford BRC"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Genes"],"abstract":"<jats:p>Pathogenic variants in the Crumbs homolog 1 (CRB1) gene lead to severe, childhood-onset retinal degeneration leading to blindness in early adulthood. There are no approved therapies, and traditional adeno-associated viral vector-based gene therapy approaches are challenged by the existence of multiple CRB1 isoforms. Here, we describe three CRB1 variants, including a novel, previously unreported variant that led to retinal degeneration. We offer a CRISPR-Cas-mediated DNA base editing strategy as a potential future therapeutic approach. This study is a retrospective case series. Clinical and genetic assessments were performed, including deep phenotyping by retinal imaging. In silico analyses were used to predict the pathogenicity of the novel variant and to determine whether the variants are amenable to DNA base editing strategies. Case 1 was a 24-year-old male with cone\u2013rod dystrophy and retinal thickening typical of CRB1 retinopathy. He had a relatively preserved central outer retinal structure and a best corrected visual acuity (BCVA) of 60 ETDRS letters in both eyes. Genetic testing revealed compound heterozygous variants in exon 9: c.2843G&gt;A, p.(Cys948Tyr) and a novel variant, c.2833G&gt;A, p.(Gly945Arg), which was predicted to likely be pathogenic by an in silico analysis. Cases 2 and 3 were two brothers, aged 20 and 24, who presented with severe cone\u2013rod dystrophy and a significant disruption of the outer nuclear layers. The BCVA was reduced to hand movements in both eyes in Case 2 and to 42 ETDRS letters in both eyes in Case 3. Case 2 was also affected with marked cystoid macular lesions, which are common in CRB1 retinopathy, but responded well to treatment with oral acetazolamide. Genetic testing revealed two c.2234C&gt;T, p.(Thr745Met) variants in both brothers. As G-to-A and C-to-T variants, all three variants are amenable to adenine base editors (ABEs) targeting the forward strand in the Case 1 variants and the reverse strand in Cases 2 and 3. Available PAM sites were detected for KKH-nSaCas9-ABE8e for the c.2843G&gt;A variant, nSaCas9-ABE8e and KKH-nSaCas9-ABE8e for the c.2833G&gt;A variant, and nSpCas9-ABE8e for the c.2234C&gt;T variant. In this case series, we report three pathogenic CRB1 variants, including a novel c.2833G&gt;A variant associated with early-onset cone\u2013rod dystrophy. We highlight the severity and rapid progression of the disease and offer ABEs as a potential future therapeutic approach for this devastating blinding condition.<\/jats:p>","DOI":"10.3390\/genes15050625","type":"journal-article","created":{"date-parts":[[2024,5,15]],"date-time":"2024-05-15T06:14:42Z","timestamp":1715753682000},"page":"625","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["In Silico CRISPR-Cas-Mediated Base Editing Strategies for Early-Onset, Severe Cone\u2013Rod Retinal Degeneration in Three Crumbs homolog 1 Patients, including the Novel Variant c.2833G&gt;A"],"prefix":"10.3390","volume":"15","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7043-3835","authenticated-orcid":false,"given":"Hoda","family":"Shamsnajafabadi","sequence":"first","affiliation":[{"name":"Nuffield Laboratory of Ophthalmology, Department of Clinical Neurosciences, Oxford University, Oxford OX3 9DU, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria","family":"Kaukonen","sequence":"additional","affiliation":[{"name":"Nuffield Laboratory of Ophthalmology, Department of Clinical Neurosciences, Oxford University, Oxford OX3 9DU, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Julia-Sophia","family":"Bellingrath","sequence":"additional","affiliation":[{"name":"Nuffield Laboratory of Ophthalmology, Department of Clinical Neurosciences, Oxford University, Oxford OX3 9DU, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert E.","family":"MacLaren","sequence":"additional","affiliation":[{"name":"Nuffield Laboratory of Ophthalmology, Department of Clinical Neurosciences, Oxford University, Oxford OX3 9DU, UK"},{"name":"Oxford Eye Hospital, Oxford University NHS Foundation Trust, John Radcliffe Hospital, Oxford OX3 9DU, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9956-6412","authenticated-orcid":false,"given":"Jasmina","family":"Cehajic-Kapetanovic","sequence":"additional","affiliation":[{"name":"Nuffield Laboratory of Ophthalmology, Department of Clinical Neurosciences, Oxford University, Oxford OX3 9DU, UK"},{"name":"Oxford Eye Hospital, Oxford University NHS Foundation Trust, John Radcliffe Hospital, Oxford OX3 9DU, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2024,5,15]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1016\/0092-8674(90)90189-L","article-title":"crumbs encodes an EGF-like protein expressed on apical membranes of Drosophila epithelial cells and required for organization of epithelia","volume":"61","author":"Tepass","year":"1990","journal-title":"Cell"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1038\/13848","article-title":"Mutations in a human homologue of Drosophila crumbs cause retinitis pigmentosa (RP12)","volume":"23","author":"Payne","year":"1999","journal-title":"Nat. Genet."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"R289","DOI":"10.1016\/j.cub.2013.03.001","article-title":"Complexities of Crumbs Function and Regulation in Tissue Morphogenesis","volume":"23","author":"Pocha","year":"2013","journal-title":"Curr. Biol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1016\/j.exer.2008.02.005","article-title":"Composition and function of the Crumbs protein complex in the mammalian retina","volume":"86","author":"Gosens","year":"2008","journal-title":"Exp. Eye Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"3328","DOI":"10.1038\/s41467-020-17009-7","article-title":"Comprehensive identification of mRNA isoforms reveals the diversity of neural cell-surface molecules with roles in retinal development and disease","volume":"11","author":"Ray","year":"2020","journal-title":"Nat. Commun."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"2659","DOI":"10.1093\/hmg\/ddl194","article-title":"Pals1\/Mpp5 is required for correct localization of Crb1 at the subapical region in polarized M\u00fcller glia cells","volume":"15","author":"Aartsen","year":"2006","journal-title":"Hum. Mol. Genet."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1016\/j.preteyeres.2014.01.001","article-title":"The CRB1 and adherens junction complex proteins in retinal development and maintenance","volume":"40","author":"Alves","year":"2014","journal-title":"Prog. Retin. Eye Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"3384","DOI":"10.1093\/hmg\/ddu048","article-title":"Targeted ablation of Crb2 in photoreceptor cells induces retinitis pigmentosa","volume":"23","author":"Alves","year":"2014","journal-title":"Hum. Mol. Genet."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1016\/j.ajo.2022.09.002","article-title":"CRB1-Associated Retinal Dystrophies: Genetics, Clinical Characteristics, and Natural History","volume":"246","author":"Georgiou","year":"2023","journal-title":"Am. J. Ophthalmol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"8654","DOI":"10.1038\/s41598-017-09035-1","article-title":"The correlation between CRB1 variants and the clinical severity of Brazilian patients with different inherited retinal dystrophy phenotypes","volume":"7","author":"Motta","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1773","DOI":"10.1016\/j.ophtha.2014.03.010","article-title":"Whole exome sequencing identifies CRB1 defect in an unusual maculopathy phenotype","volume":"121","author":"Tsang","year":"2014","journal-title":"Ophthalmology"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1001\/jamaophthalmol.2023.5886","article-title":"Optical Coherence Tomography Feature of Retinoschisis in CRB1-Associated Maculopathy","volume":"142","author":"Bellingrath","year":"2024","journal-title":"JAMA Ophthalmol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1073","DOI":"10.1093\/hmg\/ddg117","article-title":"Crumbs homolog 1 (CRB1) mutations result in a thick human retina with abnormal lamination","volume":"12","author":"Jacobson","year":"2003","journal-title":"Hum. Mol. Genet."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1086\/321263","article-title":"Leber congenital amaurosis and retinitis pigmentosa with Coats-like exudative vasculopathy are associated with mutations in the crumbs homologue 1 (CRB1) gene","volume":"69","author":"Heckenlively","year":"2001","journal-title":"Am. J. Hum. Genet."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"100699","DOI":"10.1016\/j.ajoc.2020.100699","article-title":"CRB1 related retinal degeneration with novel mutation","volume":"18","author":"Ghiam","year":"2020","journal-title":"Am. J. Ophthalmol. Case Rep."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"884","DOI":"10.1016\/j.ophtha.2017.01.047","article-title":"Genotypic and Phenotypic Characteristics of CRB1-Associated Retinal Dystrophies: A Long-Term Follow-up Study","volume":"124","author":"Talib","year":"2017","journal-title":"Ophthalmology"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2637","DOI":"10.1167\/iovs.15-18281","article-title":"Biallelic Mutations in CRB1 Underlie Autosomal Recessive Familial Foveal Retinoschisis","volume":"57","author":"Vincent","year":"2016","journal-title":"Investig. Ophthalmol. Vis. Sci."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"15","DOI":"10.1080\/13816810500481840","article-title":"A G1103R mutation in CRB1 is co-inherited with high hyperopia and Leber congenital amaurosis","volume":"27","author":"Abouzeid","year":"2006","journal-title":"Ophthalmic Genet."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1001","DOI":"10.1002\/ajmg.a.33862","article-title":"Homozygosity mapping identifies the Crumbs homologue 1 (Crb1) gene as responsible for a recessive syndrome of retinitis pigmentosa and nanophthalmos","volume":"155a","author":"Zenteno","year":"2011","journal-title":"Am. J. Med. Genet. A"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1002\/humu.21653","article-title":"CRB1 mutations in inherited retinal dystrophies","volume":"33","author":"Bujakowska","year":"2012","journal-title":"Hum. Mutat."},{"key":"ref_21","first-page":"2447","article-title":"A novel crumbs homolog 1 mutation in a family with retinitis pigmentosa, nanophthalmos, and optic disc drusen","volume":"18","author":"Paun","year":"2012","journal-title":"Mol. Vis."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"161","DOI":"10.1097\/IAE.0000000000001127","article-title":"Long-term follow-up of patients with retinitis pigmentosa type 12 caused by CRB1 Mutations: A Severe Phenotype with Considerable Interindividual Variability","volume":"37","author":"Mathijssen","year":"2017","journal-title":"Retina"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"999","DOI":"10.1089\/hum.2009.086","article-title":"Human RPE65 gene therapy for Leber congenital amaurosis: Persistence of early visual improvements and safety at 1 year","volume":"20","author":"Cideciyan","year":"2009","journal-title":"Hum. Gene Ther."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"354","DOI":"10.1038\/s41591-020-0763-1","article-title":"Initial results from a first-in-human gene therapy trial on X-linked retinitis pigmentosa caused by mutations in RPGR","volume":"26","author":"Xue","year":"2020","journal-title":"Nat. Med."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"2231","DOI":"10.1056\/NEJMoa0802268","article-title":"Effect of gene therapy on visual function in Leber\u2019s congenital amaurosis","volume":"358","author":"Bainbridge","year":"2008","journal-title":"N. Engl. J. Med."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1129","DOI":"10.1016\/S0140-6736(13)62117-0","article-title":"Retinal gene therapy in patients with choroideremia: Initial findings from a phase 1\/2 clinical trial","volume":"383","author":"MacLaren","year":"2014","journal-title":"Lancet"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"2057","DOI":"10.1038\/mt.2010.149","article-title":"Restoration of cone vision in the CNGA3-\/- mouse model of congenital complete lack of cone photoreceptor function","volume":"18","author":"Michalakis","year":"2010","journal-title":"Mol. Ther."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"50","DOI":"10.1016\/S0140-6736(17)30979-0","article-title":"Intravitreous injection of AAV2-sFLT01 in patients with advanced neovascular age-related macular degeneration: A phase 1, open-label trial","volume":"390","author":"Heier","year":"2017","journal-title":"Lancet"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"558","DOI":"10.1016\/j.ophtha.2015.10.025","article-title":"Gene Therapy for Leber Hereditary Optic Neuropathy: Initial Results","volume":"123","author":"Feuer","year":"2016","journal-title":"Ophthalmology"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1089\/humc.2013.037","article-title":"Preclinical potency and safety studies of an AAV2-mediated gene therapy vector for the treatment of MERTK associated retinitis pigmentosa","volume":"24","author":"Conlon","year":"2013","journal-title":"Hum. Gene Ther. Clin. Dev."},{"key":"ref_31","doi-asserted-by":"crossref","unstructured":"Bellingrath, J.S., McClements, M.E., Kaukonen, M., Fischer, M.D., and MacLaren, R.E. (2021). In Silico Analysis of Pathogenic CRB1 Single Nucleotide Variants and Their Amenability to Base Editing as a Potential Lead for Therapeutic Intervention. Genes, 12.","DOI":"10.3390\/genes12121908"},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"World Medical Association (2013). World Medical Association Declaration of Helsinki: Ethical Principles for Medical Research Involving Human Subjects. JAMA, 310, 2191\u20132194.","DOI":"10.1001\/jama.2013.281053"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"405","DOI":"10.1038\/gim.2015.30","article-title":"Standards and guidelines for the interpretation of sequence variants: A joint consensus recommendation of the American College of Medical Genetics and Genomics and the Association for Molecular Pathology","volume":"17","author":"Richards","year":"2015","journal-title":"Genet. Med."},{"key":"ref_34","first-page":"7.20.1","article-title":"Predicting functional effect of human missense mutations using PolyPhen-2","volume":"76","author":"Adzhubei","year":"2013","journal-title":"Curr. Protoc. Hum. Genet."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"3812","DOI":"10.1093\/nar\/gkg509","article-title":"SIFT: Predicting amino acid changes that affect protein function","volume":"31","author":"Ng","year":"2003","journal-title":"Nucleic Acids Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"W446","DOI":"10.1093\/nar\/gkab266","article-title":"MutationTaster2021","volume":"49","author":"Steinhaus","year":"2021","journal-title":"Nucleic Acids Res."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"eaao4774","DOI":"10.1126\/sciadv.aao4774","article-title":"Improved base excision repair inhibition and bacteriophage Mu Gam protein yields C: G-to-T: A base editors with higher efficiency and product purity","volume":"3","author":"Komor","year":"2017","journal-title":"Sci. Adv."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"2452","DOI":"10.1016\/j.ymthe.2022.03.023","article-title":"Reconstructed glycosylase base editors GBE2. 0 with enhanced C-to-G base editing efficiency and purity","volume":"30","author":"Sun","year":"2022","journal-title":"Mol. Ther."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"57","DOI":"10.1038\/nature26155","article-title":"Evolved Cas9 variants with broad PAM compatibility and high DNA specificity","volume":"556","author":"Hu","year":"2018","journal-title":"Nature"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1038\/s41587-020-0453-z","article-title":"Phage-assisted evolution of an adenine base editor with improved Cas domain compatibility and activity","volume":"38","author":"Richter","year":"2020","journal-title":"Nat. Biotechnol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"4333","DOI":"10.1016\/j.molcel.2021.08.008","article-title":"Engineered miniature CRISPR-Cas system for mammalian genome regulation and editing","volume":"81","author":"Xu","year":"2021","journal-title":"Mol. Cell"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Kaukonen, M., McClements, M.E., and MacLaren, R.E. (2022). CRISPR DNA Base Editing Strategies for Treating Retinitis Pigmentosa Caused by Mutations in Rhodopsin. Genes, 13.","DOI":"10.3390\/genes13081327"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"761","DOI":"10.1136\/jmedgenet-2016-103837","article-title":"Molecular findings from 537 individuals with inherited retinal disease","volume":"53","author":"Ellingford","year":"2016","journal-title":"J. Med. Genet."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"3104","DOI":"10.1093\/hmg\/ddv062","article-title":"Gene therapy into photoreceptors and M\u00fcller glial cells restores retinal structure and function in CRB1 retinitis pigmentosa mouse models","volume":"24","author":"Pellissier","year":"2015","journal-title":"Hum. Mol. Genet."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"423","DOI":"10.1016\/j.omtm.2020.12.012","article-title":"AAV-CRB2 protects against vision loss in an inducible CRB1 retinitis pigmentosa mouse model","volume":"20","author":"Buck","year":"2021","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Boon, N., Alves, C.H., Mulder, A.A., Andriessen, C.A., Buck, T.M., Quinn, P.M.J., Vos, R.M., Koster, A.J., Jost, C.R., and Wijnholds, J. (2021). Defining Phenotype, Tropism, and Retinal Gene Therapy Using Adeno-Associated Viral Vectors (AAVs) in New-Born Brown Norway Rats with a Spontaneous Mutation in Crb1. Int. J. Mol. Sci., 22.","DOI":"10.3390\/ijms22073563"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"2281","DOI":"10.1038\/nprot.2013.143","article-title":"Genome engineering using the CRISPR-Cas9 system","volume":"8","author":"Ran","year":"2013","journal-title":"Nat. Protoc."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1146\/annurev.biochem.77.061306.125255","article-title":"Mechanism of eukaryotic homologous recombination","volume":"77","author":"Sung","year":"2008","journal-title":"Annu. Rev. Biochem."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"21","DOI":"10.3389\/fgene.2018.00021","article-title":"Commentary: Programmable base editing of A\u00b7T to G\u00b7C in genomic DNA without DNA cleavage","volume":"9","author":"Matsoukas","year":"2018","journal-title":"Front. Genet."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1038\/s41587-020-0592-2","article-title":"Glycosylase base editors enable C-to-A and C-to-G base changes","volume":"39","author":"Zhao","year":"2021","journal-title":"Nat. Biotechnol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1154","DOI":"10.1038\/s41587-020-0517-0","article-title":"An engineered ScCas9 with broad PAM range and high specificity and activity","volume":"38","author":"Chatterjee","year":"2020","journal-title":"Nat. Biotechnol."},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Friedland, A.E., Baral, R., Singhal, P., Loveluck, K., Shen, S., Sanchez, M., Marco, E., Gotta, G.M., Maeder, M.L., and Kennedy, E.M. (2015). Characterization of Staphylococcus aureus Cas9: A smaller Cas9 for all-in-one adeno-associated virus delivery and paired nickase applications. Genome Biol., 16.","DOI":"10.1186\/s13059-015-0817-8"},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"14500","DOI":"10.1038\/ncomms14500","article-title":"In vivo genome editing with a small Cas9 orthologue derived from Campylobacter jejuni","volume":"8","author":"Kim","year":"2017","journal-title":"Nat. Commun."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1038\/s41586-019-1711-4","article-title":"Search-and-replace genome editing without double-strand breaks or donor DNA","volume":"576","author":"Anzalone","year":"2019","journal-title":"Nature"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"eaba5614","DOI":"10.1126\/sciadv.aba5614","article-title":"A gene therapy for inherited blindness using dCas9-VPR-mediated transcriptional activation","volume":"6","author":"Splith","year":"2020","journal-title":"Sci. Adv."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"1678","DOI":"10.1038\/s12276-023-01057-2","article-title":"Genome editing in the treatment of ocular diseases","volume":"55","author":"Choi","year":"2023","journal-title":"Exp. Mol. Med."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"839","DOI":"10.1038\/s41573-020-0084-6","article-title":"Base editing: Advances and therapeutic opportunities","volume":"19","author":"Porto","year":"2020","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_58","doi-asserted-by":"crossref","unstructured":"Guo, C., Ma, X., Gao, F., and Guo, Y. (2023). Off-target effects in CRISPR\/Cas9 gene editing. Front. Bioeng. Biotechnol., 11.","DOI":"10.3389\/fbioe.2023.1143157"},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"906","DOI":"10.1016\/j.stemcr.2019.03.002","article-title":"Human iPSC-Derived Retinas Recapitulate the Fetal CRB1 CRB2 Complex Formation and Demonstrate that Photoreceptors and M\u00fcller Glia Are Targets of AAV5","volume":"12","author":"Quinn","year":"2019","journal-title":"Stem Cell Rep."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"35","DOI":"10.1093\/hmg\/dds398","article-title":"Loss of CRB2 in the mouse retina mimics human retinitis pigmentosa due to mutations in the CRB1 gene","volume":"22","author":"Alves","year":"2013","journal-title":"Hum. Mol. Genet."},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"14009","DOI":"10.1038\/mtm.2014.9","article-title":"Specific tools for targeting and expression in M\u00fcller glial cells","volume":"1","author":"Pellissier","year":"2014","journal-title":"Mol. Ther. Methods Clin. Dev."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"3413","DOI":"10.1038\/s41596-023-00875-y","article-title":"Structure-guided AAV capsid evolution strategies for enhanced CNS gene delivery","volume":"18","author":"Gonzalez","year":"2023","journal-title":"Nat. Protoc."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"503","DOI":"10.3389\/fnins.2017.00503","article-title":"Evaluating Efficiencies of Dual AAV Approaches for Retinal Targeting","volume":"11","author":"Carvalho","year":"2017","journal-title":"Front. Neurosci."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.1016\/j.stemcr.2023.03.014","article-title":"AAV-mediated gene augmentation therapy of CRB1 patient-derived retinal organoids restores the histological and transcriptional retinal phenotype","volume":"18","author":"Boon","year":"2023","journal-title":"Stem Cell Rep."},{"key":"ref_65","doi-asserted-by":"crossref","unstructured":"Han, Z., Conley, S.M., Makkia, R., Guo, J., Cooper, M.J., and Naash, M.I. (2012). Comparative Analysis of DNA Nanoparticles and AAVs for Ocular Gene Delivery. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0052189"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"1361","DOI":"10.2147\/IJN.S157000","article-title":"DNA nanoparticles are safe and nontoxic in non-human primate eyes","volume":"13","author":"Kelley","year":"2018","journal-title":"Int. J. Nanomed."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"29","DOI":"10.1016\/bs.pmbts.2021.01.024","article-title":"CRISPR genome engineering for retinal diseases","volume":"182","author":"Kantor","year":"2021","journal-title":"Prog. Mol. Biol. Transl. Sci."},{"key":"ref_68","doi-asserted-by":"crossref","unstructured":"Elsayed, M.E.A., Kaukonen, M., Kiraly, P., Kapetanovic, J.C., and MacLaren, R.E. (2022). Potential CRISPR Base Editing Therapeutic Options in a Sorsby Fundus Dystrophy Patient. Genes, 13.","DOI":"10.3390\/genes13112103"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1089\/hum.2020.231","article-title":"Genome-Editing Strategies for Treating Human Retinal Degenerations","volume":"32","author":"Quinn","year":"2021","journal-title":"Hum. Gene Ther."},{"key":"ref_70","first-page":"387","article-title":"Bioengineering strategies for restoring vision","volume":"7","author":"Singh","year":"2023","journal-title":"Nat. Biomed. Eng."},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"1068185","DOI":"10.3389\/fnmol.2022.1068185","article-title":"Gene-agnostic therapeutic approaches for inherited retinal degenerations","volume":"15","author":"John","year":"2023","journal-title":"Front. Mol. Neurosci."}],"container-title":["Genes"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4425\/15\/5\/625\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T14:42:36Z","timestamp":1760107356000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4425\/15\/5\/625"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,15]]},"references-count":71,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2024,5]]}},"alternative-id":["genes15050625"],"URL":"https:\/\/doi.org\/10.3390\/genes15050625","relation":{},"ISSN":["2073-4425"],"issn-type":[{"value":"2073-4425","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,15]]}}}