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Retinal Degenerative Diseases XIX pp 571–576Cite as

Generation of CRB1 RP Patient-Derived iPSCs and a CRISPR/Cas9-Mediated Homology-Directed Repair Strategy for the CRB1 c.2480G>T Mutation

Part of the Advances in Experimental Medicine and Biology book series (AEMB,volume 1415)

Abstract

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.

Keywords

  • Crumbs-homologue-1 (CRB1)
  • Gene editing
  • Homology-directed repair (HDR)
  • Retinitis pigmentosa (RP)
  • iPSC-derived retinal organoids

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Acknowledgments

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.

Author information

Authors and Affiliations

  1. Department of Biomedical Engineering, Columbia University, New York, NY, USA

    Bruna Lopes da Costa & Stephen H. Tsang

  2. Edward S. Harkness Eye Institute, Department of Ophthalmology, Columbia University Irving Medical Center/New York-Presbyterian Hospital, New York, NY, USA

    Bruna Lopes da Costa, Yao Li, Sarah R. Levi, Stephen H. Tsang & Peter M. J. Quinn

  3. Jonas Children’s Vision Care, and Bernard & Shirlee Brown Glaucoma Laboratory, Department of Ophthalmology, Columbia University, New York, NY, USA

    Bruna Lopes da Costa, Yao Li, Sarah R. Levi, Stephen H. Tsang & Peter M. J. Quinn

  4. Columbia Stem Cell Initiative, Columbia University, New York, NY, USA

    Stephen H. Tsang

  5. Department of Pathology & Cell Biology, Columbia University, New York, NY, USA

    Stephen H. Tsang

  6. Institute of Human Nutrition, Columbia University, New York, NY, USA

    Stephen H. Tsang

Authors
  1. Bruna Lopes da Costa
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  2. Yao Li
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  3. Sarah R. Levi
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  4. Stephen H. Tsang
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  5. Peter M. J. Quinn
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Corresponding author

Correspondence to Peter M. J. Quinn .

Editor information

Editors and Affiliations

  1. Department of Ophthalmology, University of Pittsburgh School of Medicine, Pittsburgh, PA, USA

    John D. Ash

  2. Ocular Genomics InstituteDepartment of OphthalmologyMassachusetts Eye and Ear InfirmaryHarvard Medical School, Boston, MA, USA

    Eric Pierce

  3. Health Sciences Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK, USA

    Robert E. Anderson

  4. Department of Ophthalmology, Duke Medical Center, Durham, NC, USA

    Catherine Bowes Rickman

  5. Department of Ophthalmology, Cleveland Clinic Lerner College of Medicine, Cleveland, OH, USA

    Joe G. Hollyfield

  6. Laboratory for Retinal Cell BiologyDepartment of OphthalmologyUniversity Hospital Zurich, University of Zurich, Schlieren, Switzerland

    Christian Grimm

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© 2023 The Author(s), under exclusive license to Springer Nature Switzerland AG

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Cite this paper

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

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