CRISPR/Cas9-mediated deletion of the Wiskott-Aldrich syndrome locus causes actin cytoskeleton disorganization in murine erythroleukemia cells

18 p.-8 fig.

Detalles Bibliográficos
Autores: Fernández-Calleja, Vanessa, Fernández-Nestosa, María José, Hernandez, Pablo, Schvartzman, Jorge Bernardo, Krimer, Dora B.
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2019
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/175244
Acceso en línea:http://hdl.handle.net/10261/175244
Access Level:acceso abierto
Palabra clave:Wiskott-Aldrich
Erythroleukemia cells
Actin cytoskeleton
CRISPR/Cas9
Bruton tyrosine kinase
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spelling CRISPR/Cas9-mediated deletion of the Wiskott-Aldrich syndrome locus causes actin cytoskeleton disorganization in murine erythroleukemia cellsFernández-Calleja, VanessaFernández-Nestosa, María JoséHernandez, PabloSchvartzman, Jorge BernardoKrimer, Dora B.Wiskott-AldrichErythroleukemia cellsActin cytoskeletonCRISPR/Cas9Bruton tyrosine kinase18 p.-8 fig.Wiskott-Aldrich syndrome (WAS) is a recessive X-linked inmmunodeficiency caused by loss-of-function mutations in the gene encoding the WAS protein (WASp). WASp plays an important role in the polymerization of the actin cytoskeleton in hematopoietic cells through activation of the Arp2/3 complex. In a previous study, we found that actin cytoskeleton proteins, including WASp, were silenced in murine erythroleukemia cells defective in differentiation. Here, we designed a CRISPR/Cas9 strategy to delete a 9.5-kb genomic region encompassing the Was gene in the X chromosome of murine erythroleukemia (MEL) cells. We show that Was-deficient MEL cells have a poor organization of the actin cytoskeleton that can be recovered by restoring Was expression. We found that whereas the total amount of actin protein was similar between wild-type and Was knockout MEL cells, the latter exhibited an altered ratio of monomeric G-actin to polymeric F-actin. We also demonstrate that Was overexpression can mediate the activation of Bruton’s tyrosine kinase. Overall, these findings support the role of WASp as a key regulator of F-actin in erythroid cells.This work was supported by grants BFU2014 to Jorge B. Schvartzman, Pablo Hernández and Dora B. Krimer from the Ministerio de Economía y Competitividad of Spain, ICOOPB20224 from the National Spanish Research Council (CSIC) to Dora B Krimer and María-José Fernández-Nestosa, and PINV15-573 from CONACYT, Paraguay to María-José Fernández-Nestosa, and Jorge B Schvartzman.Peer reviewedPeerJMinisterio de Economía y Competitividad (España)Consejo Superior de Investigaciones Científicas (España)Consejo Nacional de Ciencia y Tecnología (Paraguay)Hernandez, Pablo [0000-0002-8113-1277]Krimer, Dora B. [0000-0001-9972-3994]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]201920192019info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionhttp://hdl.handle.net/10261/175244reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BFU2014-56835-Phttps://doi.org/10.7717/peerj.6284Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/1752442026-05-22T06:33:51Z
dc.title.none.fl_str_mv CRISPR/Cas9-mediated deletion of the Wiskott-Aldrich syndrome locus causes actin cytoskeleton disorganization in murine erythroleukemia cells
title CRISPR/Cas9-mediated deletion of the Wiskott-Aldrich syndrome locus causes actin cytoskeleton disorganization in murine erythroleukemia cells
spellingShingle CRISPR/Cas9-mediated deletion of the Wiskott-Aldrich syndrome locus causes actin cytoskeleton disorganization in murine erythroleukemia cells
Fernández-Calleja, Vanessa
Wiskott-Aldrich
Erythroleukemia cells
Actin cytoskeleton
CRISPR/Cas9
Bruton tyrosine kinase
title_short CRISPR/Cas9-mediated deletion of the Wiskott-Aldrich syndrome locus causes actin cytoskeleton disorganization in murine erythroleukemia cells
title_full CRISPR/Cas9-mediated deletion of the Wiskott-Aldrich syndrome locus causes actin cytoskeleton disorganization in murine erythroleukemia cells
title_fullStr CRISPR/Cas9-mediated deletion of the Wiskott-Aldrich syndrome locus causes actin cytoskeleton disorganization in murine erythroleukemia cells
title_full_unstemmed CRISPR/Cas9-mediated deletion of the Wiskott-Aldrich syndrome locus causes actin cytoskeleton disorganization in murine erythroleukemia cells
title_sort CRISPR/Cas9-mediated deletion of the Wiskott-Aldrich syndrome locus causes actin cytoskeleton disorganization in murine erythroleukemia cells
dc.creator.none.fl_str_mv Fernández-Calleja, Vanessa
Fernández-Nestosa, María José
Hernandez, Pablo
Schvartzman, Jorge Bernardo
Krimer, Dora B.
author Fernández-Calleja, Vanessa
author_facet Fernández-Calleja, Vanessa
Fernández-Nestosa, María José
Hernandez, Pablo
Schvartzman, Jorge Bernardo
Krimer, Dora B.
author_role author
author2 Fernández-Nestosa, María José
Hernandez, Pablo
Schvartzman, Jorge Bernardo
Krimer, Dora B.
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Economía y Competitividad (España)
Consejo Superior de Investigaciones Científicas (España)
Consejo Nacional de Ciencia y Tecnología (Paraguay)
Hernandez, Pablo [0000-0002-8113-1277]
Krimer, Dora B. [0000-0001-9972-3994]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv Wiskott-Aldrich
Erythroleukemia cells
Actin cytoskeleton
CRISPR/Cas9
Bruton tyrosine kinase
topic Wiskott-Aldrich
Erythroleukemia cells
Actin cytoskeleton
CRISPR/Cas9
Bruton tyrosine kinase
description 18 p.-8 fig.
publishDate 2019
dc.date.none.fl_str_mv 2019
2019
2019
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/175244
url http://hdl.handle.net/10261/175244
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BFU2014-56835-P
https://doi.org/10.7717/peerj.6284
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv PeerJ
publisher.none.fl_str_mv PeerJ
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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