Applications of genome editing technology in the targeted therapy of human diseases: mechanisms, advances and prospects

Signal Transduction and Targeted Therapy, Год журнала: 2020, Номер 5(1)

Опубликована: Янв. 3, 2020

Abstract Based on engineered or bacterial nucleases, the development of genome editing technologies has opened up possibility directly targeting and modifying genomic sequences in almost all eukaryotic cells. Genome extended our ability to elucidate contribution genetics disease by promoting creation more accurate cellular animal models pathological processes begun show extraordinary potential a variety fields, ranging from basic research applied biotechnology biomedical research. Recent progress developing programmable such as zinc-finger nucleases (ZFNs), transcription activator-like effector (TALENs) clustered regularly interspaced short palindromic repeat (CRISPR)–Cas-associated greatly expedited gene concept clinical practice. Here, we review recent advances three major (ZFNs, TALENs, CRISPR/Cas9) discuss applications their derivative reagents tools various human diseases future therapies, focusing cells models. Finally, provide an overview trials applying platforms for treatment some challenges implementation this technology.

Язык: Английский

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CRISPR/Cas9 systems: Delivery technologies and biomedical applications

Asian Journal of Pharmaceutical Sciences, Год журнала: 2023, Номер 18(6), С. 100854 - 100854

Опубликована: Окт. 21, 2023

The emergence of the clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) genome-editing system has brought about a significant revolution in realm managing human diseases, establishing animal models, and so on. To fully harness potential this potent gene-editing tool, ensuring efficient secure delivery to target site is paramount. Consequently, developing effective methods for CRISPR/Cas9 become critical area research. In review, we present comprehensive outline strategies discuss their biomedical applications system. We also provide an in-depth analysis physical, viral vector, non-viral vector strategies, including plasmid-, mRNA- protein-based approach. addition, illustrate This review highlights key factors affecting process current challenges facing system, while delineating future directions prospects that could inspire innovative strategies. aims new insights ideas advancing CRISPR/Cas9-based facilitate breakthroughs research therapeutic applications.

Язык: Английский

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Opposite Modulation of RAC1 by Mutations in TRIO Is Associated with Distinct, Domain-Specific Neurodevelopmental Disorders

The American Journal of Human Genetics, Год журнала: 2020, Номер 106(3), С. 338 - 355

Опубликована: Фев. 27, 2020

The Rho-guanine nucleotide exchange factor (RhoGEF) TRIO acts as a key regulator of neuronal migration, axonal outgrowth, axon guidance, and synaptogenesis by activating the GTPase RAC1 modulating actin cytoskeleton remodeling. Pathogenic variants in are associated with neurodevelopmental diseases, including intellectual disability (ID) autism spectrum disorders (ASD). Here, we report largest international cohort 24 individuals confirmed pathogenic missense or nonsense TRIO. mutations spread along sequence, affected show variable phenotypes. In contrast, cluster into two mutational hotspots one seventh spectrin repeat RAC1-activating GEFD1. Although all this present developmental delay neuro-behavioral phenotype, variant have more severe ID macrocephaly than do most GEFD1 variants, who display milder microcephaly. Functional studies that cause TRIO-mediated hyper- hypo-activation RAC1, respectively, observe striking correlation between activation levels head size individuals. addition, truncations vertebrate model X. tropicalis induce defects concordant human phenotype. This work demonstrates distinct clinical molecular clustering domains highlights importance tight control TRIO-RAC1 signaling development.

Язык: Английский

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Tissue-Specific Gene Inactivation inXenopus laevis: Knockout oflhx1in the Kidney with CRISPR/Cas9

Genetics, Год журнала: 2017, Номер 208(2), С. 673 - 686

Опубликована: Ноя. 30, 2017

Abstract Xenopus laevis is a classic developmental model, but its allotetraploid genome has limited our ability to perform genetic manipulations. The advance of... Studying genes involved in organogenesis often difficult because many of these are also essential for early development. frog, laevis, commonly used study processes, the presence two homeologs genes, it been use as model. Few studies have successfully CRISPR amphibians, and currently there no tissue-targeted knockout strategy described Xenopus. goal this determine whether CRISPR/Cas9-mediated gene can be targeted kidney without perturbing function. We demonstrate that targeting editing eye F0 embryos feasible. Our shows both lhx1 results disruption development function does not lead defects. Therefore, may necessary bypass defects reported upon Lhx1 protein expression or by morpholinos, antisense RNA, dominant negative constructs. establish control (slc45a2) when knocked out albinism altering This establishes feasibility tissue-specific Xenopus, providing cost-effective efficient method assessing roles implicated abnormalities amenable high-throughput drug screening techniques.

Язык: Английский

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Application of CRISPR/Cas9 gene editing technique in the study of cancer treatment

Clinical Genetics, Год журнала: 2019, Номер 97(1), С. 73 - 88

Опубликована: Июнь 24, 2019

Abstract In recent years, gene editing, especially that using clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9, has made great progress in the field of function. Rapid development editing techniques contributed to their significance medicine. Because CRISPR/Cas9 tool is not only powerful but also features such as strong specificity and high efficiency, it can accurately rapidly screen whole genome, facilitating administration therapy for specific diseases. tumor research, be used edit genomes explore mechanisms occurrence, development, metastasis. these this system been increasingly applied treatment research. treat tumors by repairing mutations or knocking out genes. To date, numerous preliminary studies have conducted on related fields. holds promise gene‐level treatment. Personalized targeted based will possibly shape future. study, we review findings research provide references future pathogenesis clinical tumors.

Язык: Английский

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Identification and functional evaluation of GRIA1 missense and truncation variants in individuals with ID: An emerging neurodevelopmental syndrome

The American Journal of Human Genetics, Год журнала: 2022, Номер 109(7), С. 1217 - 1241

Опубликована: Июнь 7, 2022

GRIA1 encodes the GluA1 subunit of α-amino-3-hydroxy-5-methyl-4-isoxazole propionate (AMPA) receptors, which are ligand-gated ion channels that act as excitatory receptors for neurotransmitter L-glutamate (Glu). AMPA (AMPARs) homo- or heteromeric protein complexes with four subunits, each encoded by different genes, to GRIA4. Although GluA1-containing AMPARs have a crucial role in brain function, human phenotype associated deleterious sequence variants has not been established. Subjects de novo missense and nonsense were identified through international collaboration. Detailed phenotypic genetic assessments subjects carried out pathogenicity was evaluated vitro characterize changes AMPAR function expression. In addition, two Xenopus gria1 CRISPR-Cas9 F0 models established vivo consequences. Seven unrelated individuals rare identified. One individual homozygous variant (p.Arg377Ter), six had heterozygous variations (p.Arg345Gln, p.Ala636Thr, p.Ile627Thr, p.Gly745Asp), p.Ala636Thr recurrent three individuals. The cohort revealed neurodevelopmental disorder mostly affecting cognition speech. Functional evaluation major subtypes carrying mutations showed profoundly perturb receptor function. stop-gain completely destroys expression AMPARs. show transient motor deficits, an intermittent seizure phenotype, significant impairment working memory mutants. These data support developmental caused both

Язык: Английский

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Xenopus Resources: Transgenic, Inbred and Mutant Animals, Training Opportunities, and Web-Based Support

Frontiers in Physiology, Год журнала: 2019, Номер 10

Опубликована: Апрель 25, 2019

Two species of the clawed frog family, Xenopus laevis and tropicalis, are widely used as tools to investigate both normal disease-state biochemistry, genetics, cell biology developmental biology. To support specialist non-specialist scientists needing access these models for their research, a number centralised resources exist around world. These include centres that hold live frozen stocks transgenic, inbred mutant animals molecular resources. This infrastructure is supported by model organism database. Here we describe much this encourage community make best use it guide resource in developing new lines libraries.

Язык: Английский

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Maximizing CRISPR/Cas9 phenotype penetrance applying predictive modeling of editing outcomes in Xenopus and zebrafish embryos

Scientific Reports, Год журнала: 2020, Номер 10(1)

Опубликована: Сен. 4, 2020

Abstract CRISPR/Cas9 genome editing has revolutionized functional genomics in vertebrates. However, edited F 0 animals too often demonstrate variable phenotypic penetrance due to the mosaic nature of outcomes after double strand break (DSB) repair. Even with high efficiency levels editing, phenotypes may be obscured by proportional presence in-frame mutations that still produce protein. Recently, studies cell culture systems have shown CRISPR/Cas9-mediated can dependent on local sequence context and predicted computational methods. Here, we similar approaches used forecast gene Xenopus tropicalis , laevis, zebrafish. We show a publicly available neural network previously trained mouse embryonic stem cultures (InDelphi-mESC) is able accurately predict early vertebrate embryos. Our observations direct implications for experiment design, allowing selection guide RNAs repair outcome signatures enriched towards frameshift mutations, maximization phenotype generation.

Язык: Английский

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Dnd1 Knockout in Sturgeons By CRISPR/Cas9 Generates Germ Cell Free Host for Surrogate Production

Animals, Год журнала: 2019, Номер 9(4), С. 174 - 174

Опубликована: Апрель 17, 2019

Sturgeons also known as living fossils are facing threats to their survival due overfishing and interference in natural habitats. Sterlet (Acipenser ruthenus) its rapid reproductive cycle small body size can be used a sterile host for surrogate production late maturing large sturgeon species. Dead end protein (dnd1) is essential migration of Primordial Germ Cells (PGCs), the origin all germ cells developing embryos. Knockout or knockdown dnd1 done order mismigrate PGCs. Previously we have MO UV aforementioned purpose, our present study CRISPR/Cas9 technology knockout dnd1. No smaller number PGCs were detected crispants, observed malformations some injected Furthermore, compared three established methods achieve sterility sterlet, found higher embryo hatching rates CRISPR/Cas9, MO, respectively.

Язык: Английский

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Applications of Genome Editing Technology in Animal Disease Modeling and Gene Therapy

Computational and Structural Biotechnology Journal, Год журнала: 2019, Номер 17, С. 689 - 698

Опубликована: Янв. 1, 2019

Genome editing technology is a technique for targeted genetic modifications, enabling the knockout and addition of specific DNA fragments. This has been widely used in various types biomedical research, clinics agriculture. In terms disease constructing appropriate animal models necessary. Combining reproductive with genome editing, many have generated basic clinical research. addition, precisely modifications allow to flourish field gene therapy. Many mutations refractory traditional therapy could be permanently corrected at level. Thus, undoubtedly promising this review, we mainly introduce applications therapies, as well its future prospects challenges.

Язык: Английский

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