A high-fidelity RNA-targeting Cas13 restores paternal Ube3a expression and improves motor functions in Angelman syndrome mice

Li, Jinhui; Shen, Zhixin; Liu, Yajing; Yan, Zixiang; Liu, Yuanhua; Lin, Xin; Tang, Junjie; Lv, Ruimin; Geng, Guannan; Xiong, Zhi‐Qi; Zhou, Chunmei; Yang, Hui

doi:10.1016/j.ymthe.2023.02.015

Cited by 17 publications

(12 citation statements)

References 40 publications

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“… 24 In contrast, RNA‐targeting CRISPR‐Cas systems, particularly the Cas13 protein family, have emerged as effective tools for transcriptome engineering, with broad applications in research and therapeutics. 14 , 17 , 25 , 26 , 27 , 28 The Cas13 proteins, guided by a single RNA molecule (gRNA), exhibit high specificity and efficiency in targeting and cleaving specific RNA transcripts. 29 Among these, RfxCas13d, or CasRx, an orthologue of CRISPR–Cas13d, has shown superior RNA knockdown efficiency and specificity compared to Cas13a and Cas13b variants.…”

Section: Discussionmentioning

confidence: 99%

“…However, it is important to note that DNA‐targeting CRISPRi systems can inadvertently silence adjacent genes within operons 24 . In contrast, RNA‐targeting CRISPR‐Cas systems, particularly the Cas13 protein family, have emerged as effective tools for transcriptome engineering, with broad applications in research and therapeutics 14,17,25–28 . The Cas13 proteins, guided by a single RNA molecule (gRNA), exhibit high specificity and efficiency in targeting and cleaving specific RNA transcripts 29 .…”

Section: Discussionmentioning

confidence: 99%

“…Preclinical studies have shown that hfCas13d can efficiently knock down disease‐causing transcripts in vivo without adverse effects. 17 …”

Section: Introductionmentioning

confidence: 99%

“…Preclinical studies have shown that hfCas13d can efficiently knock down disease-causing transcripts in vivo without adverse effects. 17 In the present study, we first explored the effects of hfCas13dmediated PLK1 knockdown on osteosarcoma cell viability, apop-…”

mentioning

confidence: 99%

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Targeted PLK1 suppression through RNA interference mediated by high‐fidelity Cas13d mitigates osteosarcoma progression via TGF‐β/Smad3 signalling

Yuan,

Cao,

Zhang

et al. 2024

J Cellular Molecular Medi

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Osteosarcoma is the most common primary bone malignancy in children and adolescents. Overexpression of polo‐like kinase 1 (PLK1) is frequent in osteosarcoma and drives disease progression and metastasis, making it a promising therapeutic target. In this study, we explored PLK1 knockdown in osteosarcoma cells using RNA interference mediated by high‐fidelity Cas13d (hfCas13d). PLK1 was found to be significantly upregulated in osteosarcoma tumour tissues compared to normal bone. sgRNA‐mediated PLK1 suppression via hfCas13d transfection inhibited osteosarcoma cell proliferation, induced G2/M cell cycle arrest, promoted apoptosis, reduced cell invasion and increased expression of the epithelial marker E‐cadherin. Proximity labelling by TurboID coupled with co‐immunoprecipitation identified novel PLK1 interactions with Smad3, a key intracellular transducer of TGF‐β signalling. PLK1 knockdown impaired Smad2/3 phosphorylation and modulated TGF‐β/Smad3 pathway inactivation. Finally, in vivo delivery of hfCas13d vectors targeting PLK1 substantially attenuated osteosarcoma xenograft growth in nude mice. Taken together, this study highlights PLK1 as a potential therapeutic target and driver of disease progression in osteosarcoma. It also demonstrates the utility of hfCas13d‐mediated gene knockdown as a strategy for targeted therapy. Further optimization of PLK1 suppression approaches may ultimately improve clinical outcomes for osteosarcoma patients.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

“…Preclinical studies have shown that hfCas13d can efficiently knock down disease‐causing transcripts in vivo without adverse effects. 17 …”

Section: Introductionmentioning

confidence: 99%

mentioning

confidence: 99%

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Targeted PLK1 suppression through RNA interference mediated by high‐fidelity Cas13d mitigates osteosarcoma progression via TGF‐β/Smad3 signalling

Yuan,

Cao,

Zhang

et al. 2024

J Cellular Molecular Medi

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“…Cas13 has also been widely used in vivo , allowing targeted RNA knockdown in xenograft tumours [ 20 , 21 ] and various tissues, including eye [ 22–24 ], ear [ 25 , 26 ], brain [ 22 , 27 , 28 ], lung [ 29–31 ], kidney [ 30 ], liver [ 30 ] and spleen [ 30 ]. Cas13 proteins, and especially Cas13d proteins, are significantly smaller than Cas9, allowing efficient package into Adeno-associated viruses (AAVs) along with its gRNA expression cassette for in vivo delivery [ 11 ].…”

Section: Crispr-cas13 As a Programmable Rnasementioning

confidence: 99%

Programmable RNA targeting with CRISPR-Cas13

Shi,

2024

RNA Biology

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The RNA-targeting CRISPR-Cas13 system has enabled precise engineering of endogenous RNAs, significantly advancing our understanding of RNA regulation and the development of RNA-based diagnostic and therapeutic applications. This review aims to provide a summary of Cas13-based RNA targeting tools and applications, discuss limitations and challenges of existing tools and suggest potential directions for further development of the RNA targeting system.

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Engineered CRISPR RNA improves the RNA cleavage efficiency of hfCas13X

Liu,

Zhang,

Wang

et al. 2024

FEBS Letters

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As the most compact variant in the Cas13 family, CRISPR‐Cas13X holds considerable promise for gene therapy applications. The development of high‐fidelity Cas13X (hfCas13X) mutants has enhanced the safety profile for in vivo applications. However, a notable reduction in on‐target cleavage efficiency accompanies the diminished collateral cleavage activity in hfCas13X. In this study, we obtained two engineered crRNA mutants that notably enhance the on‐target cleavage efficiency of hfCas13X. Furthermore, we have identified a novel crRNA structure that consistently augments the on‐target cleavage efficiency of hfCas13X across various cellular environments, without significant enhancement of its collateral activity. These findings collectively enrich the gene‐editing toolkit, presenting a more effective hfCas13X system for future research and application.

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A high-fidelity RNA-targeting Cas13 restores paternal Ube3a expression and improves motor functions in Angelman syndrome mice

Cited by 17 publications

References 40 publications

Targeted PLK1 suppression through RNA interference mediated by high‐fidelity Cas13d mitigates osteosarcoma progression via TGF‐β/Smad3 signalling

Targeted PLK1 suppression through RNA interference mediated by high‐fidelity Cas13d mitigates osteosarcoma progression via TGF‐β/Smad3 signalling

Programmable RNA targeting with CRISPR-Cas13

Engineered CRISPR RNA improves the RNA cleavage efficiency of hfCas13X

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