Project description:Rhodopsin P23H mutation is the most comment mutation causing autosomal dominant retinitis pigmentosa in the USA. The goal of this project is to compare the transcriptome changes of the Rhodopsin P23H knock-in mouse model of adRP to the wildtype control at different ages. The transcriptomic profile will help us understand the molecular events along the pathophysiology of reititis pigmentosa in this mouse model. We include the RNA seq data of Rhodopsin P23H heterozygous mouse retinas at 1, 3 and 6 months of age to compare with age-matched wildtype mouse retinas. N=3 and each sample is from an individual animal.
Project description:We report the single base pair analysis of the ocular transcriptome from wild type and BC027072 knockout animals. Comparison was analyzed to understand gene expression changes in a mouse model for early onset retinal degeneration which phenocopies a human form of autosomal recessive retinitis pigmentosa
Project description:Mutations in pre-mRNA processing factors (PRPFs) cause autosomal dominant retinitis pigmentosa (RP), but it is unclear why mutations in ubiquitously expressed genes cause retinal disease. We have generated transcriptome profiles from RP11 (PRPF31-mutated) patient-derived retinal organoids and retinal pigment epithelium (RPE), as well as Prpf31+/- mouse tissues, which revealed that disrupted alternative splicing occurred for specific splicing programmes. Mis-splicing of genes encoding pre-mRNA splicing proteins was limited to patient-specific retinal cells and Prpf31+/- mouse retinae and RPE. Mis-splicing of genes implicated in ciliogenesis and cellular adhesion was associated with severe RPE defects that include disrupted apical-basal polarity, reduced trans-epithelial resistance and phagocytic capacity, and decreased cilia length and incidence. Disrupted cilia morphology also occurred in patient-derived photoreceptors, associated with progressive degeneration and cellular stress. In situ gene-editing of a pathogenic mutation rescued protein expression and key cellular phenotypes in RPE and photoreceptors, providing proof-of-concept for future therapeutic strategies.
Project description:We report the single base pair analysis of the ocular transcriptome from wild type and BC027072 knockout animals. Comparison was analyzed to understand gene expression changes in a mouse model for early onset retinal degeneration which phenocopies a human form of autosomal recessive retinitis pigmentosa Eye mRNA profiles were generated from 3 week-old C57BL/6J and BC027072 -/- in triplicate and sequenced using the Illumina HiSeq 2500
Project description:NADPH-dependent retinol dehydrogenase 12 (RDH12) reduces all-trans-retinal in photoreceptor inner segments to regulate the phototransduction cascade and control the concentration of retinoids in the retina. Biallelic variants in RDH12 have been reported to cause autosomal recessive (AR) Leber congenital amaurosis (LCA), a severe cause of childhood blindness. Animal models have failed to recapitulate the clinical phenotype, hence, we generated human induced pluripotent stem cell (hiPSC) derived retinal organoids (RO) from the dermal fibroblasts of a female patient with homozygous missense variant c.619A>G p.(Asn207Asp) in RDH12 (RDH12-AR), and an unrelated healthy control (WT). RDH12-AR RO develop shortened rod and cone photoreceptors (from the outer limiting membrane to the tip of the outer segment), which lack RDH12 localisation in the inner segment. Bulk transcriptomic analysis revealed cone marker and apoptosis dysregulation in RDH12-AR RO compared to WT; but phospholipid transport is perturbed in RDH12-AR RO compared to an autosomal dominant hiPSC-derived RDH12-retinitis pigmentosa RO model. Our study helps to decipher the distinct disease pathways involved in RDH12 retinopathies, however, further modelling from additional patients with isogenic controls will strengthen the findings and aid identification of targets for therapeutic approaches.
Project description:To explore the mechanism associated with retinal degeneration and adeno-associated virus (AAV)-mediated gene therapy in rd10 mouse, a model of autosomal recessive retinitis pigmentosa (arRP) containing mutation of β subunit of the rod cGMP phosphodiesterase 6 (PDE6).