<i>In Vivo</i> Exon Replacement in the Mouse <i>Atp7b</i> Gene by the Cas9 System

Liu, Lili; Cao, Jiafeng; Chang, Qiurong; Xing, Fengying; Yan, Guofeng; Fu, Li; Huiyang, Wang; Ma, Zhiwei; Chen, Xuejin; Li, Yao; Li, Shangang

doi:10.1089/hum.2019.037

Cited by 4 publications

(1 citation statement)

References 34 publications

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“…Jiang et al created a single amino acid substitute rabbit model for WD, representing the most frequent WD missense mutation in Asia (p. Arg778Leu) in exon 8 of ATP7B [ 31 , 32 ]. Lately, Liu et al replaced exon 8 of the ATP7B gene in a mouse model using CRISPR/Cas9 [ 33 ]. However, a gene correction of ATP7B point mutations on human cellular level has not been described so far.…”

Section: Introductionmentioning

confidence: 99%

CRISPR/Cas9-mediated correction of mutated copper transporter ATP7B

et al. 2020

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Wilson's disease (WD) is a monogenetic liver disease that is based on a mutation of the ATP7B gene and leads to a functional deterioration in copper (Cu) excretion in the liver. The excess Cu accumulates in various organs such as the liver and brain. WD patients show clinical heterogeneity, which can range from acute or chronic liver failure to neurological symptoms. The course of the disease can be improved by a lifelong treatment with zinc or chelators such as D-penicillamine in a majority of patients, but serious side effects have been observed in a significant portion of patients, e.g. neurological deterioration and nephrotoxicity, so that a liver transplant would be inevitable. An alternative therapy option would be the genetic correction of the ATP7B gene. The novel gene therapy method CRISPR/Cas9, which has recently been used in the clinic, may represent a suitable therapeutic opportunity. In this study, we first initiated an artificial ATP7B point mutation in a human cell line using CRISPR/Cas9 gene editing, and corrected this mutation by the additional use of singlestranded oligo DNA nucleotides (ssODNs), simulating a gene correction of a WD point mutation in vitro. By the addition of 0.5 mM of Cu three days after lipofection, a high yield of CRISPR/Cas9-mediated ATP7B repaired cell clones was achieved (60%). Moreover, the repair efficiency was enhanced using ssODNs that incorporated three blocking mutations. The repaired cell clones showed a high resistance to Cu after exposure to increasing Cu concentrations. Our findings indicate that CRISPR/Cas9-mediated correction of ATP7B point mutations is feasible and may have the potential to be transferred to the clinic.

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Section: Introductionmentioning

confidence: 99%

CRISPR/Cas9-mediated correction of mutated copper transporter ATP7B

et al. 2020

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Nanotechnology-enabled gene delivery for cancer and other genetic diseases

Jiang

Gonzalez

Cordova

et al. 2023

Expert Opinion on Drug Delivery

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Navigating the CRISPR/Cas Landscape for Enhanced Diagnosis and Treatment of Wilson’s Disease

Choi,

Cha,

Kim

2024

Cells

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The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) system continues to evolve, thereby enabling more precise detection and repair of mutagenesis. The development of CRISPR/Cas-based diagnosis holds promise for high-throughput, cost-effective, and portable nucleic acid screening and genetic disease diagnosis. In addition, advancements in transportation strategies such as adeno-associated virus (AAV), lentiviral vectors, nanoparticles, and virus-like vectors (VLPs) offer synergistic insights for gene therapeutics in vivo. Wilson’s disease (WD), a copper metabolism disorder, is primarily caused by mutations in the ATPase copper transporting beta (ATP7B) gene. The condition is associated with the accumulation of copper in the body, leading to irreversible damage to various organs, including the liver, nervous system, kidneys, and eyes. However, the heterogeneous nature and individualized presentation of physical and neurological symptoms in WD patients pose significant challenges to accurate diagnosis. Furthermore, patients must consume copper-chelating medication throughout their lifetime. Herein, we provide a detailed description of WD and review the application of novel CRISPR-based strategies for its diagnosis and treatment, along with the challenges that need to be overcome.

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In Vivo Exon Replacement in the Mouse Atp7b Gene by the Cas9 System

Cited by 4 publications

References 34 publications

CRISPR/Cas9-mediated correction of mutated copper transporter ATP7B

CRISPR/Cas9-mediated correction of mutated copper transporter ATP7B

Nanotechnology-enabled gene delivery for cancer and other genetic diseases

Navigating the CRISPR/Cas Landscape for Enhanced Diagnosis and Treatment of Wilson’s Disease

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