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Published on: May 25, 2018
Allele-specific gene editing to rescue dominant CRX-associated LCA7 phenotypes in a retinal organoid model
Kathleen R Chirco1, Shereen Chew2, Anthony T Moore3
1Department of Ophthalmology, University of California San Francisco, San Francisco, CA, USA; The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California San Francisco, San Francisco, CA, USA; Divison of Neuroscience, Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, OR, USA; Casey Eye Institute, Oregon Health & Science University, Portland, OR, USA.
Abstract:
Cases of Leber congenital amaurosis caused by mutations in CRX (LCA7) exhibit an early form of the disease and show signs of significant photoreceptor dysfunction and eventual loss. To establish a translational in vitro model system to study gene-editing-based therapies, we generated LCA7 retinal organoids harboring a dominant disease-causing mutation in CRX. Our LCA7 retinal organoids develop signs of immature and dysfunctional photoreceptor cells, providing us with a reliable in vitro model to recapitulate LCA7. Furthermore, we performed a proof-of-concept study in which we utilize allele-specific CRISPR/Cas9-based gene editing to knock out mutant CRX and saw moderate rescue of photoreceptor phenotypes in our organoids. This work provides early evidence for an effective approach to treat LCA7, which can be applied more broadly to other dominant genetic diseases.
Insights
Researchers developed a novel in vitro model for Leber congenital amaurosis type 7 (LCA7) using retinal organoids. Gene editing partially restored photoreceptor function, offering hope for treating this inherited retinal disease.
Area of Science:
- Ophthalmology
- Genetics
- Molecular Biology
Background:
- Leber congenital amaurosis type 7 (LCA7) is an inherited retinal disease caused by mutations in the CRX gene.
- LCA7 leads to early-onset vision loss due to photoreceptor dysfunction and degeneration.
Purpose of the Study:
- To create a translational in vitro model for LCA7 using retinal organoids.
- To test the efficacy of allele-specific CRISPR/Cas9 gene editing for treating LCA7.
Main Methods:
- Generation of LCA7 retinal organoids with a dominant CRX mutation.
- Utilizing allele-specific CRISPR/Cas9 gene editing to target and knock out the mutant CRX allele.
- Assessing photoreceptor development and function in the organoid model.
Main Results:
- LCA7 retinal organoids exhibited immature and dysfunctional photoreceptor cells, successfully modeling the disease.
- CRISPR/Cas9 gene editing demonstrated a moderate rescue of photoreceptor phenotypes in the organoids.
- Proof-of-concept achieved for gene editing as a therapeutic strategy for LCA7.
Conclusions:
- The developed LCA7 retinal organoid model is a reliable platform for studying LCA7 and testing therapies.
- Allele-specific gene editing shows potential for treating LCA7 by correcting the underlying genetic defect.
- This approach may be applicable to other dominant genetic retinal diseases.
