RT info:eu-repo/semantics/article
T1 CRISPR/Cas9-mediated deletion of the Wiskott-Aldrich syndrome locus causes actin cytoskeleton disorganization in murine erythroleukemia cells
A1 Fernández Calleja, Vanessa
A1 Fernández de Nestosa, María José
A1 Hernández, Pablo
A1 Schvartzman, Jorge Bernardo
A1 Krimer, Dora B.
A2 Universidad Nacional de Asunción. Facultad Politécnica
K1 Hematology
K1 Molecular Biology
AB 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.
PB PeerJ, Inc.
YR 2019
FD 2019-01-16
LK http://hdl.handle.net/20.500.14066/3848
UL http://hdl.handle.net/20.500.14066/3848
LA eng
NO Fernández Calleja, V., Fernández Nestosa, M. J., Hernández, P., Schvartzman, J. B., & Krimer, D. B. (2019). CRISPR/Cas9-mediated deletion of the Wiskott-Aldrich syndrome locus causes actin cytoskeleton disorganization in murine erythroleukemia cells. PeerJ, Artículo 7:e6284.https://doi.org/10.7717/peerj.6284
NO Corresponding author: Dora B. Krimer, dbkrimer@cib.csic.es.
NO Consejo Nacional de Ciencia y Tecnología
DS MINDS@UW
RD 03-sep-2026