Clustered Regularly Interspaced Short Palindromic Repeats-Based Genome Surgery for the Treatment of Autosomal Dominant Retinitis Pigmentosa

Abstract

As a proof of concept, our results suggest that the ablate-and-replace strategy can ameliorate disease progression as measured by photoreceptor structure and function for both of the human mutation knock-in models. These results demonstrate the potency of the ablate-and-replace strategy to treat RP caused by different Rho mutations. Furthermore, because ablate-and-replace treatment is mutation independent, this strategy may be used to treat a wide array of dominant diseases in ophthalmology and other fields. Clinical trials using ablate-and-replace gene therapy would allow researchers to determine if this strategy provides any benefits for patients with diseases of interest.

Mutation-Dependent Pathomechanisms Determine the Phenotype in the Bestrophinopathies

“…Although these patients may experience an initial improvement of their condition, in the long-term progression of the degenerative disease with corresponding functional limitations would be expected as multifocal subretinal deposits and abnormal autofluorescence in the ARB phenotype may further develop from ER stress-induced accumulation of cytosolic aggregation-prone proteins. Based on our model, it may be more appropriate for these patients to follow a so-called "ablate-and-replace" strategy for full recovery of vision as suggested by Tsai et al [53].…”

Mutationsabhängige Mechanismen und deren Bedeutung für zielgerichtete Behandlungsstrategien am Beispiel von Bestrophin 1 und den Bestrophinopathien

“…"Ablate-and-Replace"-Strategie für eine vollständige Wiederherstellung der Sehfähigkeit besser geeignet sein. In einer kürzlich veröffentlichen Studie konnte gezeigt werden, dass mittels einer vorgeschalteten Geninaktivierung durch Verwendung der CRISPR/Cas-Technologie in Kombination mit einer Genersatztherapie der pathologische Phänotyp in einem Mausmodell für Retinitis pigmentosa nachhaltig verbessert werden konnte [36].…”

Single AAV-mediated scarless genome editing in dysfunctional retinal neurons mediates robust visual restoration in mice

“…4 Recently, the CRISPR-Cas9-mediated allele knock-out genome editing strategy, based on non-homologous end joining (NHEJ) has been successfully applied to correct gain-of-function mutations via AAV. 5, 6, 7, 8 One of the unique advantages of the genome editing approach is that it allows local treatment of the genome, such that the approach does not depend on the size of the target gene. However, genome editing for loss-of-function mutations in larger genes that require local replacement of the mutated sequence with a wildtype counterpart (mutation replacement) has not been successful in treatment of neuronal disorders primarily affecting neurons, due to its low editing efficiency.…”

“…3 CRISPR-Cas9-mediated allele knock-out based on error-prone non-homologous end joining (NHEJ) can correct gain-of-function mutations with AAV. 4–7 However, many loss-of-function mutations in larger genes still require local replacement with a wildtype sequence (scarless genome editing).…”