CRISPR-assisted transcription activation by phase-separation proteins

Protein & Cell, Год журнала: 2023, Номер 14(12), С. 874 - 887

Опубликована: Март 11, 2023

The clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 system has been widely used for genome engineering and transcriptional regulation in many different organisms. Current CRISPR-activation (CRISPRa) platforms often require multiple components because of inefficient activation. Here, we fused phase-separation proteins to dCas9-VPR (dCas9-VP64-P65-RTA) observed robust increases activation efficiency. Notably, human NUP98 (nucleoporin 98) FUS (fused sarcoma) IDR domains were best at enhancing activity, with dCas9-VPR-FUS (VPRF) outperforming the other CRISPRa systems tested this study both efficiency simplicity. dCas9-VPRF overcomes target strand bias widens gRNA designing windows without affecting off-target effect dCas9-VPR. These findings demonstrate feasibility using assist gene expression support broad appeal basic clinical applications.

Язык: Английский

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Advancements in the synergy of isothermal amplification and CRISPR-cas technologies for pathogen detection

Frontiers in Bioengineering and Biotechnology, Год журнала: 2023, Номер 11

Опубликована: Окт. 10, 2023

In the realm of pathogen detection, isothermal amplification technology has emerged as a swift, precise, and sensitive alternative to conventional PCR. This paper explores fundamental principles recombinase polymerase (RPA) recombinase-aid (RAA) reviews current status integrating CRISPR-Cas system with RPA/RAA techniques. Furthermore, this confluence technology, providing comprehensive review enhancements existing combined methodologies such SHERLOCK DETECTR. We investigate practical applications in conjunction for highlighting how integrated approach significantly advances both research clinical implementation field. aims provide readers concise understanding fusion offering insights into their utility, ongoing enhancements, promising prospects detection.

Язык: Английский

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Toward biomanufacturing of next-generation bacterial nanocellulose (BNC)-based materials with tailored properties: A review on genetic engineering approaches

Biotechnology Advances, Год журнала: 2024, Номер 74, С. 108390 - 108390

Опубликована: Май 31, 2024

Язык: Английский

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CRISPR-Cas9 mediated understanding of plants’ abiotic stress-responsive genes to combat changing climatic patterns

Functional & Integrative Genomics, Год журнала: 2024, Номер 24(4)

Опубликована: Июль 30, 2024

Язык: Английский

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Engineering CRISPR/Cas9 therapeutics for cancer precision medicine

Frontiers in Genetics, Год журнала: 2024, Номер 15

Опубликована: Апрель 25, 2024

The discovery of Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated protein 9 (Cas9) technology has revolutionized field cancer treatment. This review explores usage CRISPR/Cas9 for editing investigating genes involved in human carcinogenesis. It provides insights into the development CRISPR as a genetic tool. Also, it recent developments tools available designing systems targeting oncogenic Further, we delve an overview biology, highlighting key alterations signaling pathways whose deletion prevents malignancies. fundamental knowledge enables deeper understanding how can be tailored to address specific aberrations offer personalized therapeutic approaches. In this review, showcase studies preclinical trials that show utility disrupting targets, modulating tumor microenvironment increasing efficiency anti treatments. also insight use high throughput screens biomarker identifications based screening drug discovery. conclusion, offers exciting engineering therapeutics treatment highlights transformative potential innovation effective

Язык: Английский

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Progress and pitfalls of gene editing technology in CAR-T cell therapy: a state-of-the-art review

Frontiers in Oncology, Год журнала: 2024, Номер 14

Опубликована: Июнь 7, 2024

CAR-T cell therapy has shown remarkable promise in treating B-cell malignancies, which sparked optimism about its potential to treat other types of cancer as well. Nevertheless, the Expectations solid tumors and non-B hematologic malignancies have not been met. Furthermore, safety concerns regarding use viral vectors current personalized production process are bottlenecks that limit widespread use. In recent years gene editing technology opened a new way unleash latent potentials lessen associated challenges. Moreover, tools paved manufacturing cells fully non-viral approach well providing universal, off-the-shelf product. Despite all advantages strategies, off-target activity classical (ZFNs, TALENs, CRISPR/Cas9) remains major concern. Accordingly, several efforts made reduce their genotoxicity, leading introduction advanced with an improved profile. this review, we begin by examining tools, overview how these technologies currently being applied clinical trials therapies. Following this, explore various strategies aimed at enhancing efficacy therapy.

Язык: Английский

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Advancing crop disease resistance through genome editing: a promising approach for enhancing agricultural production

Frontiers in Genome Editing, Год журнала: 2024, Номер 6

Опубликована: Июнь 26, 2024

Modern agriculture has encountered several challenges in achieving constant yield stability especially due to disease outbreaks and lack of long-term disease-resistant crop cultivars. In the past, economically important crops had a major impact on food security economy. On other hand climate-driven emergence new pathovars or changes their host specificity further poses serious threat sustainable agriculture. At present, chemical-based control strategies are frequently used microbial pathogens pests, but they have detrimental environment also resulted development resistant phyto-pathogens. As replacement, cultivating engineered can help minimize negative regular pesticides environment. Although traditional breeding genetic engineering been instrumental improvement certain limitations such as labour intensity, time consumption, low efficiency. this regard, genome editing emerged one potential tools for improving resistance by targeting multiple traits with more accuracy For instance, techniques, CRISPR/Cas9, CRISPR/Cas13, base editing, TALENs, ZFNs, meganucleases, proved successful through targeted mutagenesis, gene knockouts, knockdowns, modifications, activation target genes. CRISPR/Cas9 is unique among these techniques because its remarkable efficacy, risk off-target repercussions, ease use. Some primary targets developing CRISPR-mediated host-susceptibility genes (the S method), (R genes) pathogen material that prevents development, broad-spectrum resistance. The use methods notably ameliorate transform agricultural practices future. This review highlights phyto-pathogens productivity. Next, we discussed while focusing editing. We provided an update accomplishments improve against bacterial, fungal viral different systems. Finally, highlighted future systems enhancing

Язык: Английский

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Recent updates on allogeneic CAR-T cells in hematological malignancies

Cancer Cell International, Год журнала: 2024, Номер 24(1)

Опубликована: Сен. 3, 2024

CAR-T cell therapy is known as an effective in patients with hematological malignancies. Since 2017, several autologous (auto-CAR-T) drugs have been approved by the US Food and Drug Administration (FDA) for treatment of some kinds relapsed/refractory However, fail to respond these due high manufacturing time, batch-to-batch variation, poor quality insufficient quantity primary T cells, their expansion function. cells prepared from allogeneic sources (allo-CAR-Ts) can be alternative option overcome obstacles. Recently, allo-CAR-Ts entered into early clinical trials. Despite promising preclinical results, there are two main barriers, including graft-versus-host disease (GvHD) allo-rejection that may decline safety efficacy clinic. The successful development products depends on starter source, gene editing method, ability escape immune rejection prevent GvHD. Here, we summarize technologies potential various developing highlight advantages We also describe data focusing allo-CAR-T blood malignancies discuss challenges future perspectives therapeutic applications.

Язык: Английский

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CRISPR-Cas9 enrichment and long read sequencing for fine mapping in plants

Plant Methods, Год журнала: 2020, Номер 16(1)

Опубликована: Сен. 1, 2020

Genomic methods for identifying causative variants trait loci applicable to a wide range of germplasm are required plant biologists and breeders understand the genetic control variation.

Язык: Английский

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Targeted epigenetic repression by CRISPR/dSaCas9 suppresses pathogenic DUX4-fl expression in FSHD

Molecular Therapy — Methods & Clinical Development, Год журнала: 2020, Номер 20, С. 298 - 311

Опубликована: Дек. 10, 2020

Facioscapulohumeral muscular dystrophy (FSHD) is caused by incomplete silencing of the disease locus, leading to pathogenic misexpression DUX4 in skeletal muscle. Previously, we showed that CRISPR inhibition could successfully target and repress FSHD myocytes. However, an effective therapy will require both efficient delivery therapeutic components muscles long-term repression locus. Thus, re-engineered our platform allow vivo more potent epigenetic repressors. We designed FSHD-optimized regulatory cassette drive muscle-specific expression dCas9 from Staphylococcus aureus fused HP1α, HP1γ, MeCP2 transcriptional domain, or SUV39H1 SET domain. Targeting each regulator promoter/exon 1 increased chromatin at specifically suppressing its genes myocytes a mouse model disease. Importantly, minimizing using smaller Cas9 ortholog allowed cassettes be effectively packaged into adeno-associated virus (AAV) vectors for delivery. By engineering compatible with AAV gene therapy, have laid groundwork clinical use dCas9-based effectors muscle disorders.

Язык: Английский

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