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http://hdl.handle.net/2445/148117
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DC Field | Value | Language |
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dc.contributor.author | Calatayud Aristoy, Carles | - |
dc.contributor.author | Carola, Giulia | - |
dc.contributor.author | Fernandez-Carasa, Irene | - |
dc.contributor.author | Valtorta, Marco | - |
dc.contributor.author | Jiménez-Delgado, Senda | - |
dc.contributor.author | Díaz, Mònica | - |
dc.contributor.author | Soriano i Fradera, Jordi | - |
dc.contributor.author | Cappelletti, Graziella | - |
dc.contributor.author | García-Sancho, Javier | - |
dc.contributor.author | Raya Chamorro, Ángel | - |
dc.contributor.author | Consiglio, Antonella | - |
dc.date.accessioned | 2020-01-17T13:03:27Z | - |
dc.date.available | 2020-01-17T13:03:27Z | - |
dc.date.issued | 2019-05-02 | - |
dc.identifier.issn | 2045-2322 | - |
dc.identifier.uri | http://hdl.handle.net/2445/148117 | - |
dc.description.abstract | Patient-specific induced pluripotent stem cells (iPSCs) are a powerful tool to investigate the molecular mechanisms underlying Parkinson's disease (PD), and might provide novel platforms for systematic drug screening. Several strategies have been developed to generate iPSC-derived tyrosine hydroxylase (TH)-positive dopaminergic neurons (DAn), the clinically relevant cell type in PD; however, they often result in mixed neuronal cultures containing only a small proportion of TH-positive DAn. To overcome this limitation, we used CRISPR/Cas9-based editing to generate a human iPSC line expressing a fluorescent protein (mOrange) knocked-in at the last exon of the TH locus. After differentiation of the TH-mOrange reporter iPSC line, we confirmed that mOrange expression faithfully mimicked endogenous TH expression in iPSC-derived DAn. We also employed calcium imaging techniques to determine the intrinsic functional differences between dopaminergic and non-dopaminergic ventral midbrain neurons. Crucially, the brightness of mOrange allowed direct visualization of TH-expressing cells in heterogeneous cultures, and enabled us to isolate live mOrange-positive cells through fluorescence-activated cell sorting, for further differentiation. This technique, coupled to refined imaging and data processing tools, could advance the investigation of PD pathogenesis and might offer a platform to test potential new therapeutics for PD and other neurodegenerative diseases. | - |
dc.format.extent | 9 p. | - |
dc.format.mimetype | application/pdf | - |
dc.language.iso | eng | - |
dc.publisher | Nature Publishing Group | - |
dc.relation.isformatof | Reproducció del document publicat a: https://doi.org/10.1038/s41598-019-43080-2 | - |
dc.relation.ispartof | Scientific Reports, 2019, vol. 9, p. 6811 | - |
dc.relation.uri | https://doi.org/10.1038/s41598-019-43080-2 | - |
dc.rights | cc-by (c) Calatayud Aristoy, Carles et al., 2019 | - |
dc.rights.uri | http://creativecommons.org/licenses/by/3.0/es | - |
dc.source | Articles publicats en revistes (Patologia i Terapèutica Experimental) | - |
dc.subject.classification | Malaltia de Parkinson | - |
dc.subject.classification | Ciències de la salut | - |
dc.subject.classification | Enginyeria genètica | - |
dc.subject.other | Parkinson's disease | - |
dc.subject.other | Medical sciences | - |
dc.subject.other | Genetic engineering | - |
dc.title | CRISPR/Cas9-mediated generation of a tyrosine hydroxylase reporter iPSC line for live imaging and isolation of dopaminergic neurons | - |
dc.type | info:eu-repo/semantics/article | - |
dc.type | info:eu-repo/semantics/publishedVersion | - |
dc.identifier.idgrec | 690555 | - |
dc.date.updated | 2020-01-17T13:03:28Z | - |
dc.relation.projectID | info:eu-repo/grantAgreement/EC/FP7/311736/EU//PD-HUMMODEL | - |
dc.relation.projectID | info:eu-repo/grantAgreement/EC/H2020/713140/EU//MESO_BRAIN | - |
dc.rights.accessRights | info:eu-repo/semantics/openAccess | - |
dc.identifier.pmid | 31048719 | - |
Appears in Collections: | Articles publicats en revistes (Patologia i Terapèutica Experimental) Articles publicats en revistes (Física de la Matèria Condensada) Articles publicats en revistes (Institut d'lnvestigació Biomèdica de Bellvitge (IDIBELL)) |
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690555.pdf | 3.13 MB | Adobe PDF | View/Open |
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