Mutations in the Crumbs-homologue-1 (CRB1) gene lead to a spectrum of severe inherited retinal diseases, including retinitis pigmentosa (RP). The establishment of a genotype–phenotype correlation in CRB1 patients has been difficult due to the substantial variability and phenotypic overlap between CRB1-associated diseases. This phenotypic modulation may be due to several factors, including genetic modifiers, deep intronic mutations, isoform diversity, and copy number variations. Induced pluripotent stem cell (iPSC)-derived patient retinal organoids are novel tools that can provide sensitive, quantitative, and scalable phenotypic assays. CRB1 RP patient iPSC-derived retinal organoids have shown reproducible phenotypes compared to healthy retinal organoids. However, having genetically defined iPSC isogenic controls that take into account potential phenotypic modulation is crucial. In this study, we generated iPSC from an early-onset CRB1 patient and developed a correction strategy for the c.2480G>T, p.(Gly827Val) CRB1 mutation using CRISPR/Cas9-mediated homology-directed repair.
- Crumbs-homologue-1 (CRB1)
- Gene editing
- Homology-directed repair (HDR)
- Retinitis pigmentosa (RP)
- iPSC-derived retinal organoids
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S.H.T. and The Jonas Children’s Vision Care and Bernard & Shirlee Brown Glaucoma Laboratory are supported by the National Institutes of Health (P30EY019007, R01EY018213, R01EY024698, R01EY026682, R24EY027285, U01EY030580), National Cancer Institute Core (5P30CA013696), Foundation Fighting Blindness (TA-NMT-0116-0692-COLU), the Research to Prevent Blindness (RPB) Physician-Scientist Award. B.L.D. is a recipient of the Capes PhD scholarship. P.M.J.Q. is the current recipient of a Curing Retinal Blindness Foundation (CRBF) grant, a Knights Templar Eye Foundation (KTEF) Career Starter grant, an Uplifting Athletes Young Investigator grant, and a New York Stem Cell Foundation (NYSCF)—Druckenmiller Fellowship.
Conflict of Interest
Stephen H. Tsang receives financial support from Abeona Therapeutics, Inc. and Emendo. He is also the founder of Rejuvitas and is on the scientific and clinical advisory board for Nanoscope Therapeutics.
Editors and Affiliations
© 2023 The Author(s), under exclusive license to Springer Nature Switzerland AG
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da Costa, B.L., Li, Y., Levi, S.R., Tsang, S.H., Quinn, P.M.J. (2023). Generation of CRB1 RP Patient-Derived iPSCs and a CRISPR/Cas9-Mediated Homology-Directed Repair Strategy for the CRB1 c.2480G>T Mutation. In: Ash, J.D., Pierce, E., Anderson, R.E., Bowes Rickman, C., Hollyfield, J.G., Grimm, C. (eds) Retinal Degenerative Diseases XIX. Advances in Experimental Medicine and Biology, vol 1415. Springer, Cham. https://doi.org/10.1007/978-3-031-27681-1_83
Publisher Name: Springer, Cham
Print ISBN: 978-3-031-27680-4
Online ISBN: 978-3-031-27681-1