Optimization of HITI-Mediated Gene Insertion for Rhodopsin and Peripherin-2 in Mouse Rod Photoreceptors: Targeting Dominant Retinitis Pigmentosa

Investigative Ophthalmology & Visual Science, Journal Year: 2024, Volume and Issue: 65(13), P. 38 - 38

Published: Nov. 18, 2024

Purpose: Among the genome-editing methods for repairing disease-causing mutations resulting in autosomal dominant retinitis pigmentosa, homology-independent targeted integration (HITI)-mediated gene insertion of normal form causative is useful because it allows development mutation-agnostic therapeutic products. In this study, we aimed rapid optimization and validation HITI-treatment constructs approach developing various target genes mouse models retinal degeneration. Methods: We constructed Cas9-driven HITI cassettes plasmid vectors to treat Rho gene. A workflow utilizing vivo electroporation was established validate efficacy these constructs. Single-cell genotyping conducted evaluate allelic donor insertion. The potency adeno-associated virus (AAV) examined by section immunohistochemistry optomotor response (OMR) Rho+/P23H mutant mice. also Prph2 examine workflow. Results: optimized achieved 80% 90% transduced rod photoreceptor cells. This construct effectively suppressed degeneration induced visual restoration Rhodopsin demonstrated AAV are adaptable locus. Conclusions: study showcases a highly effective against dominant-negative inheritance inherited dystrophy. These findings suggest potential utility genes, advancing therapy products diverse genetic disorders.

Language: Английский

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G protein-coupled receptors in cochlea: Potential therapeutic targets for hearing loss

Frontiers in Molecular Neuroscience, Journal Year: 2022, Volume and Issue: 15

Published: Oct. 12, 2022

The prevalence of hearing loss-related diseases caused by different factors is increasing worldwide year year. Currently, however, the patient’s loss has not been effectively improved. Therefore, there an urgent need to adopt new treatment measures and techniques help improve therapeutic effect loss. G protein-coupled receptors (GPCRs), as crucial cell surface receptors, can widely participate in physiological pathological processes, particularly play essential role many disease occurrences be served promising targets. However, no specific drugs on market have found target GPCRs cochlea. Interestingly, recent studies demonstrated that various pathogenic process related cochlea including heredity, noise, ototoxic drugs, cochlear structure, so on. In this review, we comprehensively summarize functions 53 known their relationships with loss, highlight advances used study cryo-EM, AI, GPCR drug screening, gene therapy vectors, CRISPR editing technology, well discuss depth future direction novel GPCR-based development for Collectively, review facilitate basic (pre-) clinical research area, provide beneficial emerging therapies.

Language: Английский

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Genetic, molecular and biochemical basis of the auditory aging: lessons from experimental models

Auditio, Journal Year: 2022, Volume and Issue: 6

Published: Jan. 1, 2022

Age-related hearing loss (ARHL) affects one in three people older than 65 years and is the most prevalent sensorineural deficit. This type of precedes accelerates onset cognitive impairment associated with an increased risk for neurodegenerative diseases such as dementia Alzheimer disease. The progression ARHL influenced by genetic factors, which are still poorly understood, environmental particular include exposure to excessive noise ototoxic substances. At present, no effective drug treatments available prevention or treatment, therefore research this field a priority. In field, animal models offer crucial tool i) identifying new genes ARHL, ii) understanding cellular molecular basis auditory ageing iii) defining therapeutic targets evaluating candidate treatments.

Language: Английский

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Efficient workflow for validating homology-independent targeted integration-mediated gene insertion in rod photoreceptor cells to treat dominant-negative mutations causing retinitis pigmentosa

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Nov. 9, 2023

Abstract Among the genome-editing methods for repairing disease-causing mutations resulting in dominant inhibition, homology-independent targeted integration (HITI)-mediated gene insertion of normal form causative is useful because it allows development mutation-agnostic therapeutic products. For rapid optimization and validation highly effective HITI-treatment constructs against dominant-negative inheritance inherited retinal dystrophy, we improved available both plasmid adeno-associated virus (AAV) vectors, established a workflow that uses vivo electroporation to verify efficacy. By targeting mouse Rhodopsin gene, derived construct which HITI-mediated occurs 80%-90% transduced rod photoreceptor cells. This suppressed degeneration induced visual restoration mutant mice. The rhodopsin were shown be AAV this construction Peripherin 2 gene. These findings suggest reported here may generation various target genes therapy

Language: Английский

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