Immunogold Labeling to Detect Streptococcus pyogenes Cas9 in Cell Culture and Tissues by Electron Microscopy
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Otros documentos de la autoría: Ulloa-Navas, María José; García-Tárraga, Patricia; García-Verdugo, Jose Manuel; Herranz-Pérez, Vicente
Metadatos
Mostrar el registro completo del ítemcomunitat-uji-handle:10234/9
comunitat-uji-handle2:10234/36080
comunitat-uji-handle3:10234/36082
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INVESTIGACIONMetadatos
Título
Immunogold Labeling to Detect Streptococcus pyogenes Cas9 in Cell Culture and Tissues by Electron MicroscopyAutoría
Fecha de publicación
2019Editor
Mary Ann LiebertISSN
2573-1599; 2573-1602Cita bibliográfica
María José Ulloa-Navas, Patricia García-Tárraga, José Manuel García-Verdugo, and Vicente Herranz-Pérez. The CRISPR Journal. Dec 2019 2:6. 395-405. http://doi.org/10.1089/crispr.2019.0032Tipo de documento
info:eu-repo/semantics/articleVersión de la editorial
https://www.liebertpub.com/doi/full/10.1089/crispr.2019.0032Versión
info:eu-repo/semantics/submittedVersionPalabras clave / Materias
Resumen
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 ... [+]
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 oligodendrocyte precursor cells, Cas9 was found in cytoplasmic vesicles. In Cas9 constitutively expressing oligodendrocyte precursors, the enzyme was located in the cytoplasm of nondividing cells. Finally, while in the liver Cas9 was detected in different cell types, in the brain we found no specifically labeled cells. In conclusion, immunoelectron microscopy opens a new spectrum of opportunities to study the CRISPR-Cas9 system in a more precise manner. [-]
Descripción
Final publication is available from Mary Ann Liebert, Inc., publishers https://doi.org/10.1089/crispr.2019.0032.
Publicado en
The CRISPR Journal, 2019, vol. 2, no 6Proyecto de investigación
Nano-scaffolding for neuronal migration and generation project - Spanish Ministry of Science, Innovation and Universities: PCI2018-093062; Red de Terapia Celular: TerCel-RD16/0011/0026; McDonald Fellowship from the Multiple Sclerosis International FederationDerechos de acceso
Copyright © Mary Ann Liebert, Inc., publishers
http://rightsstatements.org/vocab/InC/1.0/
info:eu-repo/semantics/openAccess
http://rightsstatements.org/vocab/InC/1.0/
info:eu-repo/semantics/openAccess
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