Specific knockdown of Htra2 by CRISPR-CasRx prevents acquired sensorineural hearing loss in mice

Guo, Yang; Lei, Han; Han, Shuang; Tang, Honghai; Wang, Shengyi; Cui, Chong; Chen, Bing; Li, Huawei; Shu, Yilai

doi:10.1016/j.omtn.2022.04.014

Cited by 13 publications

(11 citation statements)

References 49 publications

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“…Because of their intrinsic RNase activity [36][37][38][39][40][41][42] , their high programmability, and their ability to be used in mammalian cells to knock down the expression of a target gene 36,41,[47][48][49]73 , RNAtargeting CRISPR effector proteins are a potentially powerful class of agents for gene silencing.…”

Section: Discussionmentioning

confidence: 99%

“…To target the mATXN2 mRNA, we used the Cas13d nuclease from Ruminococcus flavefaciens XPD3002 (RfxCas13d) 41 , a programmable RNA-targeting effector protein that we 47 and others 48,49 have demonstrated can silence target gene expression in the nervous system. To facilitate the identification of crRNAs for mATXN2, we created a reporter plasmid carrying the mATXN2 protein-coding sequence fused to an enhanced green fluorescence protein (EGFP) via a self-cleaving T2A peptide, linking mATXN2 expression to EGFP fluorescence.…”

Section: Targeting Ataxin-2 With Rfxcas13dmentioning

confidence: 99%

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Mitigating a TDP-43 proteinopathy by targeting ataxin-2 using RNA-targeting CRISPR effector proteins

Moore

Powell

et al. 2023

Preprint

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The TDP-43 proteinopathies, which include amyotrophic lateral sclerosis and frontotemporal dementia, are a devastating group of neurodegenerative disorders that are characterized by the mislocalization and aggregation of TDP-43. Here we demonstrate that RNA-targeting CRISPR effector proteins, a programmable class of gene silencing agents that includes the Cas13 family of enzymes and Cas7-11, can be used to mitigate TDP-43 pathology when programmed to target ataxin-2, a modifier of TDP-43-associated toxicity. In addition to inhibiting the aggregation and transit of TDP-43 to stress granules, we find that the in vivo delivery of an ataxin-2-targeting Cas13 system to a mouse model of TDP-43 proteinopathy improved functional deficits, extended survival, and reduced the severity of neuropathological hallmarks. Further, we benchmark RNA-targeting CRISPR platforms against ataxin-2 and find that high-fidelity forms of Cas13 possess improved transcriptome-wide specificity compared to Cas7-11 and a first-generation effector. Our results demonstrate the potential of CRISPR technology for TDP-43 proteinopathies.

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Section: Discussionmentioning

confidence: 99%

Section: Targeting Ataxin-2 With Rfxcas13dmentioning

confidence: 99%

Mitigating a TDP-43 proteinopathy by targeting ataxin-2 using RNA-targeting CRISPR effector proteins

Moore

Powell

et al. 2023

Preprint

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“…Additionally, applying mini Cas‐proteins to deliver fused effectors by AAV and effectively targeting endogenous RNA in vivo underscores the potential these platforms possess to direct diverse functions in complex systems. Moreover, as the Li and Shu labs have also previously demonstrated Cas13d–mediated RNA knockdown in the context of other forms of hearing loss, this remains a promising emerging area of application for preclinical programmable RNA‐targeting development [77] …”

Section: Programmable Rbps In Preclinical Applicationsmentioning

confidence: 98%

Engineered RNA‐Binding Proteins: Studying and Controlling RNA Regulation

Sinnott,

Cao,

Dickinson

2024

Israel Journal of Chemistry

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The complexity of eukaryotic organisms is intricately tied to transcriptome‐level processes, notably alternative splicing and the precise modulation of gene expression through a sophisticated interplay involving RNA‐binding protein (RBP) networks and their RNA targets. Recent advances in our understanding of the molecular pathways responsible for this control have paved the way for the development of tools capable of steering and managing RNA regulation and gene expression. The fusion between a rapidly developing understanding of endogenous RNA regulation and the burgeoning capabilities of CRISPR‐Cas and other programmable RBP platforms has given rise to an exciting frontier in engineered RNA regulators. This review offers an overview of the existing toolkit for constructing synthetic RNA regulators using programmable RBPs and effector domains, capable of altering RNA sequence composition or fate, and explores their diverse applications in both basic research and therapeutic contexts.

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“…Unlike the CRISPR-Cas9 system targeting DNA, CRISPR-Cas13 system edits disease-associated RNA transcripts, which is transient and potentially reversible, thus also offering improved safety. Two recent studies have revealed the potential of the CRISPR-Cas13 system in gene therapy for the repair of hearing loss caused by mutations, including CRISPR-Cas13X ( Xiao et al, 2022 ) and CRISPR-CasRx ( Guo et al, 2022 ).…”

Section: Emerging G Protein-coupled Receptor-based Treatmentmentioning

confidence: 99%

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

Guo²,

Fu³

et al. 2022

Front. Mol. Neurosci.

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

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Specific knockdown of Htra2 by CRISPR-CasRx prevents acquired sensorineural hearing loss in mice

Cited by 13 publications

References 49 publications

Mitigating a TDP-43 proteinopathy by targeting ataxin-2 using RNA-targeting CRISPR effector proteins

Mitigating a TDP-43 proteinopathy by targeting ataxin-2 using RNA-targeting CRISPR effector proteins

Engineered RNA‐Binding Proteins: Studying and Controlling RNA Regulation

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

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