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dc.contributor.authorPetazzi, Paolo
dc.contributor.authorTorres-Ruiz Raul, Raul 
dc.contributor.authorFidanza, Antonella
dc.contributor.authorRoca-Ho, Heleia
dc.contributor.authorGutierrez-Agüera, Francisco
dc.contributor.authorCastaño, Julio
dc.contributor.authorRodriguez Perales, Sandra 
dc.contributor.authorDíaz de la Guardia, Rafael
dc.contributor.authorLópez-Millán, Belén
dc.contributor.authorBigas, Anna
dc.contributor.authorForrester, Lesley M
dc.contributor.authorBueno, Clara
dc.contributor.authorMenéndez, Pablo
dc.date.accessioned2020-03-20T18:08:48Z
dc.date.available2020-03-20T18:08:48Z
dc.date.issued2020-02-27
dc.identifier.citationMol Ther Nucleic Acids. 2020;20:196-204.es_ES
dc.identifier.issn2162-2531es_ES
dc.identifier.urihttp://hdl.handle.net/20.500.12105/9296
dc.description.abstractHuman pluripotent stem cells (hPSCs) and mesenchymal stromal/stem cells (hMSCs) are clinically relevant sources for cellular therapies and for modeling human development and disease. Many stem cell-based applications rely on the ability to activate several endogenous genes simultaneously to modify cell fate. However, genetic intervention of these cells remains challenging. Several catalytically dead Cas9 (dCas9) proteins fused to distinct activation domains can modulate gene expression when directed to their regulatory regions by a specific single-guide RNA (sgRNA). In this study, we have compared the ability of the first-generation dCas9-VP64 activator and the second-generation systems, dCas9-SAM and dCas9-SunTag, to induce gene expression in hPSCs and hMSCs. Several stem cell lines were tested for single and multiplexed gene activation. When the activation of several genes was compared, all three systems induced specific and potent gene expression in both single and multiplexed settings, but the dCas9-SAM and dCas9-SunTag systems resulted in the highest and most consistent level of gene expression. Simultaneous targeting of the same gene with multiple sgRNAs did not result in additive levels of gene expression in hPSCs nor hMSCs. We demonstrate the robustness and specificity of second-generation dCas9 activators as tools to simultaneously activate several endogenous genes in clinically relevant human stem cells.es_ES
dc.description.sponsorshipWe thank CERCA/Generalitat de Catalunya and Fundació Josep Carreras-Obra Social la Caixa for their institutional support. We thank Jose Luis Sardina (IJC, Barcelona) for technical assistance with the teratoma assays. Financial support for this work was obtained from the Catalunya Goverment (SGR330 and PERIS 2017-2019), the Spanish Ministry of Economy and Competitiveness (SAF2016-80481-R), the European Research Council (CoG-2014-646903), and the Fundación Leo Messi to P.M.; the Spanish Association against Cancer (AECC-CI-2015) and the Health Institute Carlos III (ISCIII/FEDER, PI17/01028) to C.B.; the Biotechnology and Biological Sciences Research Council (BBRSC) to L.M.F. and A.F.; and the Spanish National Research and Development Plan (ISCIII/FEDER, PI17/02303) and the AEI/MICIU EXPLORA Project (BIO2017-91272-EXP) to S.R.-P. P.M. is an investigator of the Spanish Cell Therapy Cooperative Network (TERCEL). R.T.-R. is supported by a postdoctoral fellowship from the Asociación Española Contra el Cáncer (AECC).es_ES
dc.language.isoenges_ES
dc.publisherElsevier es_ES
dc.type.hasVersionVoRes_ES
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.subjectCRISPRes_ES
dc.subjectSAMes_ES
dc.subjectSunTages_ES
dc.subjectdCas9 activatorses_ES
dc.subjecthMSCses_ES
dc.subjecthPSCses_ES
dc.titleRobustness of Catalytically Dead Cas9 Activators in Human Pluripotent and Mesenchymal Stem Cellses_ES
dc.typejournal articlees_ES
dc.rights.licenseAtribución-NoComercial-CompartirIgual 4.0 Internacional*
dc.identifier.pubmedID32171171es_ES
dc.format.volume20es_ES
dc.format.page196-204es_ES
dc.identifier.doi10.1016/j.omtn.2020.02.009es_ES
dc.contributor.funderGovernment of Catalonia (España) 
dc.contributor.funderFundación Josep Carreras Contra la Leucemia 
dc.contributor.funderFundación La Caixa 
dc.contributor.funderMinisterio de Economía y Competitividad (España) 
dc.contributor.funderInstituto de Salud Carlos III 
dc.contributor.funderAsociación Española Contra el Cáncer 
dc.description.peerreviewedes_ES
dc.relation.publisherversionhttps://doi.org/10.1016/j.omtn.2020.02.009.es_ES
dc.identifier.journalMolecular therapy. Nucleic acidses_ES
dc.repisalud.institucionCNIOes_ES
dc.repisalud.orgCNIOCNIO::Unidades técnicas::Unidad de Citogenética Moleculares_ES
dc.relation.projectIDinfo:eu_repo/grantAgreement/ES/sAF2016-80481-Res_ES
dc.relation.projectIDinfo:eu_repo/grantAgreement/ES/PI17/01028es_ES
dc.relation.projectIDinfo:eu_repo/grantAgreement/ES/PI17/02303es_ES
dc.rights.accessRightsopen accesses_ES


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Atribución-NoComercial-CompartirIgual 4.0 Internacional
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