Modeling Dominant and Recessive Forms of Retinitis Pigmentosa by Editing Three Rhodopsin-Encoding Genes in Xenopus Laevis Using Crispr/Cas9

Feehan, Joanna M.; Chiu, Colette N.; Stanar, Paloma; Tam, Beatrice M.; Ahmed, Sheikh N.; Moritz, Orson L.

doi:10.1038/s41598-017-07153-4

Cited by 27 publications

(28 citation statements)

References 67 publications

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“…For example, in the case of rhodopsin adRP, any CRISPR approach must be allele specific or it would convert an adRP phenotype to a more severe arRP one, and these studies have not been done on human rhodopsin genomic sequences in the context of one mutant and one WT allele. Furthermore, a recent study of CRISPR/Cas9 targeting in fertilised Xenopus eggs showed that in-frame deletions frequently caused dominant retinal degeneration associated with rhodopsin biosynthesis defects, while frameshift phenotypes were consistent with knockout (Feehan et al, 2017). Given that NHEJ is random, there is almost a 1:3 chance that CRISPR gene editing could replace one dominant rhodopsin mutation with another dominant mutation within individual rods, even if the editing were allele specific.…”

Section: Therapeutic Approaches To Rhodopsin Rpmentioning

confidence: 99%

The molecular and cellular basis of rhodopsin retinitis pigmentosa reveals potential strategies for therapy

Athanasiou¹,

Aguilà²,

Bellingham³

et al. 2018

Progress in Retinal and Eye Research

255

328

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Inherited mutations in the rod visual pigment, rhodopsin, cause the degenerative blinding condition, retinitis pigmentosa (RP). Over 150 different mutations in rhodopsin have been identified and, collectively, they are the most common cause of autosomal dominant RP (adRP). Mutations in rhodopsin are also associated with dominant congenital stationary night blindness (adCSNB) and, less frequently, recessive RP (arRP). Recessive RP is usually associated with loss of rhodopsin function, whereas the dominant conditions are a consequence of gain of function and/or dominant negative activity. The in-depth characterisation of many rhodopsin mutations has revealed that there are distinct consequences on the protein structure and function associated with different mutations. Here we categorise rhodopsin mutations into seven discrete classes; with defects ranging from misfolding and disruption of proteostasis, through mislocalisation and disrupted intracellular traffic to instability and altered function. Rhodopsin adRP offers a unique paradigm to understand how disturbances in photoreceptor homeostasis can lead to neuronal cell death. Furthermore, a wide range of therapies have been tested in rhodopsin RP, from gene therapy and gene editing to pharmacological interventions. The understanding of the disease mechanisms associated with rhodopsin RP and the development of targeted therapies offer the potential of treatment for this currently untreatable neurodegeneration.

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Section: Therapeutic Approaches To Rhodopsin Rpmentioning

confidence: 99%

The molecular and cellular basis of rhodopsin retinitis pigmentosa reveals potential strategies for therapy

Athanasiou¹,

Aguilà²,

Bellingham³

et al. 2018

Progress in Retinal and Eye Research

255

328

View full text Add to dashboard Cite

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“…In vitro fertilization and microinjections were performed at 18°C as previously described (Feehan et al, 2017). Eggs and sperm were incubated in a petri dish for 20 min, the follicle cell sheath was removed from fertilized embryos using 2% cysteine and gentle shaking, and then the embryos were tightly packed into a monolayer in 2% agarose injection plates flooded with 0.4x MMR + 6% Ficoll.…”

Section: Microinjections Embryo Selection and Tadpole Rearingmentioning

confidence: 99%

Distinct roles for prominin-1 and photoreceptor cadherin in outer segment disc morphogenesis in CRISPR-altered X. laevis

Carr

Stanar

Moritz

2020

Journal of Cell Science

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Mutations in prominin-1 (prom1) and photoreceptor cadherin (cdhr1) are associated with inherited retinal degenerative disorders but their functions remain unknown. Here, we used CRISPR-Cas9 to generate prom1-null, cdhr1-null, and prom1 plus cdhr1 double-null Xenopuslaevis and then documented the effects of these mutations on photoreceptor structure and function. Prom1-null mutations resulted in severely dysmorphic photoreceptors comprising overgrown and disorganized disc membranes. Cone outer segments were more severely affected than rods and had an impaired electroretinogram response. Cdhr1-null photoreceptors did not appear grossly dysmorphic, but ultrastructural analysis revealed that some disc membranes were overgrown or oriented vertically within the plasma membrane. Double-null mutants did not differ significantly from prom1-null mutants. Our results indicate that neither prom1 nor cdhr1 are necessary for outer segment disc membrane evagination or the fusion event that controls disc sealing. Rather, they are necessary for the higher-order organization of the outer segment. Prom1 may align and reinforce interactions between nascent disc leading edges, a function more critical in cones for structural support. Cdhr1 may secure discs in a horizontal orientation prior to fusion and regulate cone lamellae size.This article has an associated First Person interview with the first author of the paper.

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“…Xenopus embryos have long been used to study fundamental aspects of early eye development and are used to model diseases such as microphthalmia, retinitis pigmentosa, exudative vitreoretinopathy, and aniridia. Feehan et al (2017) utilized CRISPR/Cas9-mediated mutations in Xenopus genes encoding rhodopsin to model both dominant and recessive forms of retinitis pigmentosa, a disease caused by retinal degeneration that leads to gradual loss of sight. Assays on retinal extracts and confocal microscopy were used to characterize the genotype-phenotype relationships.…”

Section: The Landscape Of Human Disease Research In Xenopusmentioning

confidence: 99%

Xenbase: Facilitating the Use of Xenopus to Model Human Disease

Fisher

James-Zorn

et al. 2019

Front. Physiol.

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At a fundamental level most genes, signaling pathways, biological functions and organ systems are highly conserved between man and all vertebrate species. Leveraging this conservation, researchers are increasingly using the experimental advantages of the amphibian Xenopus to model human disease. The online Xenopus resource, Xenbase, enables human disease modeling by curating the Xenopus literature published in PubMed and integrating these Xenopus data with orthologous human genes, anatomy, and more recently with links to the Online Mendelian Inheritance in Man resource (OMIM) and the Human Disease Ontology (DO). Here we review how Xenbase supports disease modeling and report on a meta-analysis of the published Xenopus research providing an overview of the different types of diseases being modeled in Xenopus and the variety of experimental approaches being used. Text mining of over 50,000 Xenopus research articles imported into Xenbase from PubMed identified approximately 1,000 putative disease- modeling articles. These articles were manually assessed and annotated with disease ontologies, which were then used to classify papers based on disease type. We found that Xenopus is being used to study a diverse array of disease with three main experimental approaches: cell-free egg extracts to study fundamental aspects of cellular and molecular biology, oocytes to study ion transport and channel physiology and embryo experiments focused on congenital diseases. We integrated these data into Xenbase Disease Pages to allow easy navigation to disease information on external databases. Results of this analysis will equip Xenopus researchers with a suite of experimental approaches available to model or dissect a pathological process. Ideally clinicians and basic researchers will use this information to foster collaborations necessary to interrogate the development and treatment of human diseases.

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Modeling Dominant and Recessive Forms of Retinitis Pigmentosa by Editing Three Rhodopsin-Encoding Genes in Xenopus Laevis Using Crispr/Cas9

Cited by 27 publications

References 67 publications

The molecular and cellular basis of rhodopsin retinitis pigmentosa reveals potential strategies for therapy

The molecular and cellular basis of rhodopsin retinitis pigmentosa reveals potential strategies for therapy

Distinct roles for prominin-1 and photoreceptor cadherin in outer segment disc morphogenesis in CRISPR-altered X. laevis

Xenbase: Facilitating the Use of Xenopus to Model Human Disease

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