All-in-one AAV-mediated Nrl gene inactivation rescues retinal degeneration in Pde6a mice

Retinitis pigmentosa (RP) is a complex group of inherited retinal diseases characterized by progressive death of photoreceptor cells and eventual blindness. Pde6a, which encodes a cGMP-specific phosphodiesterase, is a crucial pathogenic gene for autosomal recessive RP (RP43); there is no effective t...

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Bibliographic Details
Published in:JCI insight 2024-12, Vol.9 (24)
Main Authors: Liu, Zhiquan, Chen, Siyu, Lo, Chien-Hui, Wang, Qing, Sun, Yang
Format: Article
Language:English
Subjects:
Animals
Basic-Leucine Zipper Transcription Factors
CRISPR-Cas Systems - genetics
Cyclic Nucleotide Phosphodiesterases, Type 6 - genetics
Dependovirus - genetics
Disease Models, Animal
Eye Proteins - genetics
Eye Proteins - metabolism
Gene Editing - methods
Genetic Therapy - methods
Humans
Mice
Mutation
Retina - metabolism
Retina - pathology
Retinal Degeneration - genetics
Retinal Degeneration - pathology
Retinal Degeneration - therapy
Retinitis Pigmentosa - genetics
Retinitis Pigmentosa - pathology
Retinitis Pigmentosa - therapy
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Summary:Retinitis pigmentosa (RP) is a complex group of inherited retinal diseases characterized by progressive death of photoreceptor cells and eventual blindness. Pde6a, which encodes a cGMP-specific phosphodiesterase, is a crucial pathogenic gene for autosomal recessive RP (RP43); there is no effective therapy for this form of RP. The compact CRISPR/Staphylococcus aureus Cas9 (CRISPR/SaCas9) system, which can be packaged into a single adeno-associated virus (AAV), holds promise for simplifying effective gene therapy. Here, we demonstrated that all-in-one AAV-SaCas9-mediated Nrl gene inactivation can efficiently prevent retinal degeneration in a RP mouse model with Pde6anmf363/nmf363 mutation. We screened single-guide RNAs capable of efficiently editing the mouse Nrl gene in N2a cells and then achieved effective gene editing by using a single AAV to codeliver SaCas9 and an optimal Nrl-sg2 into the mouse retina. Excitingly, in vivo inactivation of Nrl improved photoreceptor cell survival and rescued retinal function in treated Pde6a-deficient mice. Thus, we showed that a practical, gene-independent method, AAV-SaCas9-mediated Nrl inactivation, holds promise for future therapeutic applications in patients with RP.
ISSN:2379-3708
2379-3708
DOI:10.1172/jci.insight.178159