Search results for "CRISPR-Cas9"
showing 8 items of 8 documents
Inactivation of peroxisomal ABC transporters, ABCD1 and ABCD2 in BV-2 microglial cells : Towards a better understanding of X-linked adrenoleukodystro…
2018
X-linked adrenoleukodystrophy (X-ALD) is a severe neurodegenerative disorder characterized by very-long-chain fatty acid (VLCFA) accumulation resulting from a peroxisomal β-oxidation defect. The disease is caused by mutations in the ABCD1 gene, which encodes for a peroxisomal half ABC transporter predicted, like its closest homologue ABCD2, to participate in the entry of VLCFA-CoA into the peroxisome, the unique site of their β-oxidation. Progress in understanding the physiopathogenesis of X-ALD suffers from the lack of appropriate cell and animal models. Since peroxisomal defects in microglia seem to be a key element of the onset of the disease, we generated four microglial cell lines unab…
Immunogold Labeling to Detect Streptococcus pyogenes Cas9 in Cell Culture and Tissues by Electron Microscopy
2019
Final publication is available from Mary Ann Liebert, Inc., publishers https://doi.org/10.1089/crispr.2019.0032. The CRISPR-Cas9 system is a powerful and yet precise DNA-editing tool in rapid development. By combining immunogold labeling and electron microscopy with the novel CRISPR-Cas9 system, we propose a new method to gain insight into the biology of this tool. In this study, we analyzed different Cas9-induced systems such as HEK293T cell line, murine oligodendrocyte progenitor cells, brain and liver to detect Cas9 expression by immunoelectron microscopy. Our results show that while Cas9 expression could be found in the nuclei and nucleopores of transfected HEK293T cells, in transfected…
XPO1E571K Mutation Modifies Exportin 1 Localisation and Interactome in B-cell Lymphoma
2020
The XPO1 gene encodes exportin 1 (XPO1) that controls the nuclear export of cargo proteins and RNAs. Almost 25% of primary mediastinal B-cell lymphoma (PMBL) and classical Hodgkin lymphoma (cHL) cases harboured a recurrent XPO1 point mutation (NM_003400, chr2:g61718472C>
Proposal of a Genome Editing System for Genetic Resistance to Tomato Spotted Wilt Virus
2014
Viruses provoke considerable economical losses in agriculture. New molecular approaches to develop genetic resistance based on translational genomics and precision genetic modifications are highly expected. The type II Clustered, Regularly Interspaced Palindromic Repeats (CRISPR) system including Cas9 nuclease represent a promising and very powerful tool to specifically modulate the expression and activity of genes involved in biotic stress responses. In this study, we describe an approach to develop a platform system based on CRISPR system for genome editing technology in tomato. Tomato is an excellent plant for this approach considering the high-quality genome sequence, the rapid life cyc…
Deploying Genome Editing Tools for Dissecting the Biology of Nut Trees
2019
Walnuts are among the most important nut crops grown in temperate regions of the world. Commercial production in California, and increasingly worldwide, relies on only few clonally grafted scion genotypes, particularly “Chandler,” and more recently clonally propagated disease-resistant rootstocks. Diseases, nematodes, insect pests, abiotic stresses, and other nutritional and environmental factors, can reduce walnut productivity and quality, affecting grower profitability. The California Walnut Breeding Program at UC Davis has developed and released scion cultivars and rootstocks to help address some of these problems. Sequencing of the walnut genome is expected to speed walnut breeding by f…
Chi dice che i batteri fanno male? I CRISPR ed il loro potenziale genetico. In Le scienze naturali nella scuola
2019
L’immaginario collettivo non ha un’opinione buona e nemmeno benevola sui batteri. Sono spesso legati a concetti di mancanza di igiene, malattie ed a volte anche a situazioni più gravi financo la morte. È indubbio che alcuni generi di procarioti sono agenti patogeni di specifiche malattie infettive che, in passato più che oggi, hanno mietuto vittime nel genere umano. Ma che dire dei batteri lattici degli yogurt o della nostra flora batterica intestinale? C’è da aggiungere anche una lunga serie di organismi procarioti, non patogeni, che hanno, negli ultimi due secoli, costituito sistemi biologici idonei allo studio della biologia molecolare e della genetica molecolare. In definitiva è molto p…
Algunas consideraciones bioéticas y jurídicas acerca de la Declaración de Ginebra sobre edición del genoma humano hereditario a la vista de las Decla…
2021
[ES] La reciente Declaración de Ginebra sobre edición del genoma humano denuncia la posición central que han ocupado tanto la Ciencia como la Bioética hasta la fecha, afirmando que ambas mantienen una posición decididamente favorable respecto a esta bioingeniería, y todo lo cual pone de manifiesto la acuciante necesidad de reorientar el debate hacia el campo del Derecho. El presente artículo tiene por objeto analizar pormenorizadamente las consideraciones vertidas en este documento, formulando interrogantes sobre algunas de ellas, y poniendo en valor la conveniencia de articular un consenso multidisciplinar encabezado por los derechos humanos, en particular, de aquellos grupos más vulnerabl…
Desarrollo de modelos experimentales de LGMDD2 y rastreo de fármacos
2023
La distrofia muscular de cinturas tipo D2 (Limb-girdle muscular dystrophy type D2, LGMDD2) es una enfermedad genética ultra rara para la cual no existe tratamiento. Las principales características clínicas de la LGMDD2 son la debilidad y degeneración muscular progresiva que afecta a las cinturas pélvica y escapular de forma predominante. En 2013 se identificó la LGMDD2 como una miopatía autosómica dominante causada por una mutación en el gen de la transportina 3 (TNPO3), dando como resultado una proteína mutante con una extensión de 15 aminoácidos adicionales: TNPO3mut. TNPO3 es una β-importina responsable de la importación nuclear de proteínas ricas en serina/arginina (proteínas SR), invol…