Браузер не даёт сохранять данные (например, приватный режим): закладки пропадут после закрытия вкладки.
Поиск статей PubMed
Ключевые слова, автор, журнал, годы — результаты из PubMed, аннотации с переводом на русский, закладки в браузере.
Найдено: 26 081
Страница 108 из 1000-
Non-surgical In Vivo Germ Cell-mediated Gene Editing by CRISPR Mutagenic Chain Reaction with the Aid of Magnetic Nanoparticles.
Gene therapy via germline cells leads to a permanent genetic modification. The promise of this method is due to its potential for providing a stable therapeutic effect for all who inherit the gene of interest. If germinal therapy is successfully performed, it can eliminate certain diseases from the family and the population. The feasibility of genetic modification in the human germline raises several controversial and bioethical issues. However, gene transfer via male and female germinal cells has been recently explored in animal models. Previous studies have shown that delivering DNA to the testes followed by electroporation is relatively successful in producing germline-mediated alterations. Since this method includes surgical procedures, non-surgical, safer, and less timeconsuming methods would be ideal. Herein, we discuss a potential approach for nonsurgical in vivo germ cell-mediated gene editing by CRISPR mutagenic chain reaction with the aid of magnetic nanoparticles.
-
Cancer CRISPR Screens In Vivo.
Clustered regularly interspaced short palindromic repeats (CRISPR) screening is a powerful toolset for investigating diverse biological processes. Most CRISPR screens to date have been performed with in vitro cultures or cellular transplant models. To interrogate cancer in animal models that more closely recapitulate the human disease, autochthonous direct in vivo CRISPR screens have recently been developed that can identify causative drivers in the native tissue microenvironment. By empowering multiplexed mutagenesis in fully immunocompetent animals, direct in vivo CRISPR screens enable the rapid generation of patient-specific avatars that can guide precision medicine. This Opinion article discusses the current status of in vivo CRISPR screens in cancer and offers perspectives on future applications.
-
[Application of CRISPR/Cas9 mediated gene editing in trees].
The CRISPR/Cas9 system, which can induce precise modifications at a target gene, has been recognized as the most promising gene editing technology, and has played an important role in precision crop breeding. It also provides a new strategy for fundamental researches and molecular breeding of forest trees. Recently, CRISPR/Cas9-mediated gene editing has been applied more extensively in tree genetic studies. It has not only succeeded in developing new drought- resistance or disease-resistant cultivars, but also shows a great potential in regulating lignin biosynthesis and shortening the breeding cycle of forest trees. In this review, we summarize the application and advance of CRISPR/Cas9 in gene function identification and genetic improvement of forest plants. We also discuss the related problems and future perspectives. This review aims to provide a useful reference for tree functional genomics and genetic engineering breeding.
-
Questioning the proposed translational pathway for germline genome editing.
-
Systematic discovery of CRISPR-boosted CAR T cell immunotherapies.
Chimeric antigen receptor (CAR) T cell therapy has shown remarkable success in treating blood cancers, but CAR T cell dysfunction remains a common cause of treatment failure1. Here we present CELLFIE, a CRISPR screening platform for enhancing CAR T cells across multiple clinical objectives. We performed genome-wide screens in human primary CAR T cells, with readouts capturing key aspects of T cell biology, including proliferation, target cell recognition, activation, apoptosis and fratricide, and exhaustion. Screening hits were prioritized using a new in vivo CROP-seq2 method in a xenograft model of human leukaemia, establishing several gene knockouts that boost CAR T cell efficacy. Most notably, we discovered that RHOG knockout is a potent and unexpected CAR T cell enhancer, both individually and together with FAS knockout, which was validated across multiple in vivo models, CAR designs and sample donors, and in patient-derived cells. Demonstrating the versatility of the CELLFIE platform, we also conducted combinatorial CRISPR screens to identify synergistic gene pairs and saturation base-editing screens to characterize RHOG variants. In summary, we discovered, validated and biologically characterized CRISPR-boosted CAR T cells that outperform standard CAR T cells in widely used benchmarks, establishing a foundational resource for optimizing cell-based immunotherapies.
-
Generation of novel self-incompatible Brassica napus by CRISPR/Cas9.
-
Improved Genome Editing via Oviductal Nucleic Acids Delivery (i-GONAD): Protocol Steps and Additional Notes.
The clustered regularly interspaced short palindromic repeats (CRISPR) technology has made it possible to produce genome-edited (GE) animals more easily and rapidly than before. In most cases, GE mice are produced by microinjection (MI) or by in vitro electroporation (EP) of CRISPR reagents into fertilized eggs (zygotes). Both of these approaches require ex vivo handling of isolated embryos and their subsequent transfer into another set of mice (called recipient or pseudopregnant mice). Such experiments are performed by highly skilled technicians (especially for MI). We recently developed a novel genome editing method, called "GONAD (Genome-editing via Oviductal Nucleic Acids Delivery)," which can completely eliminate the ex vivo handling of embryos. We also made improvements to the GONAD method, termed "improved-GONAD (i-GONAD)." The i-GONAD method involves injection of CRISPR reagents into the oviduct of an anesthetized pregnant female using a mouthpiece-controlled glass micropipette under a dissecting microscope, followed by EP of the entire oviduct allowing the CRISPR reagents to enter into the zygotes present inside the oviduct, in situ. After the i-GONAD procedure, the mouse recovered from anesthesia is allowed to continue the pregnancy to full term to deliver its pups. The i-GONAD method does not require pseudopregnant female animals for embryo transfer, unlike the methods relying on ex vivo handling of zygotes. Therefore, the i-GONAD method can reduce the number of animals used, compared to the traditional methods. In this chapter, we describe some newer technical tips about the i-GONAD method. Additionally, even though the detailed protocols of GONAD and i-GONAD have been published elsewhere (Gurumurthy et al., Curr Protoc Hum Genet 88:15.8.1-15.8.12, 2016 Nat Protoc 14:2452-2482, 2019), we provide all the protocol steps of i-GONAD in this chapter so that the reader can find most of the information, needed for performing i-GONAD experiments, in one place.
-
Ordo-Responsibility for Germline Gene Editing.
The case of twins born with genes modified by He Jiankui highlights the need for international governance of germline gene editing (GGE). This article proposes a global framework that utilizes "ordo-responsibilities." This is a pragmatic ethical approach open to pluralism and grounded in principles of human dignity and human rights. Ordo-responsibility is pragmatic in (1) accepting generally available values on a global level (e.g., human dignity, human rights) and (2) seeking achievable implementation. Genetic science is practiced globally in ways that transcend borders. As such, its practice must take account of the vast complexity of cultural, ethical, legal, and anthropological convictions. Here, we explain the basic structure of an appropriate rule-finding process, outline a possible pathway toward an international framework, and discuss minimal requirements that are needed in that endeavor. We thereby contribute to the debate on how to govern genome-editing technologies and GGE globally.
-
Cas14: Big Advances from Small CRISPR Proteins.
-
Co-expression of Cas9 and single-guided RNAs in Escherichia coli streamlines production of Cas9 ribonucleoproteins.
CRISPR/Cas9 ribonucleoprotein (RNP) complexes are promising biological tools with diverse biomedical applications. However, to date there are no efficient methods that can produce these proteins at large scales and low cost. Here, we present a streamlined method for direct production of Cas9 RNPs from Escherichia coli by co-expression of Cas9 and the target-specific single-guided RNAs. Harnessing an ultrahigh-affinity CL7/Im7 purification system recently developed we achieve one-step purification of the self-assembling CRISPR/Cas RNPs, including the commonly used Cas9 and Cas12a, within half a day and with a ~fourfold higher yield than incumbent methods. The prepared Cas RNPs show remarkable stability in the absence of RNase inhibitors, as well as profound gene-editing efficiency in vitro and in vivo. Our method is convenient, cost-effective, and can be used to prepare other CRISPR/Cas RNPs.
PubMed отдаёт не больше 10 000 первых результатов — уточните запрос, чтобы увидеть остальные.