dc.contributor.author | Bonafont, Jose | |
dc.contributor.author | Mencía, Ángeles | |
dc.contributor.author | García, Marta | |
dc.contributor.author | Torres-Ruiz, Raul | |
dc.contributor.author | Rodriguez Perales, Sandra | |
dc.contributor.author | Carretero, Marta | |
dc.contributor.author | Chacón-Solano, Esteban | |
dc.contributor.author | Modamio-Høybjør, Silvia | |
dc.contributor.author | Marinas, Lucía | |
dc.contributor.author | León, Carlos | |
dc.contributor.author | Escamez, María J | |
dc.contributor.author | Hausser, Ingrid | |
dc.contributor.author | Del Río, Marcela | |
dc.contributor.author | Murillas, Rodolfo | |
dc.contributor.author | Larcher, Fernando | |
dc.date.accessioned | 2019-05-20T10:00:37Z | |
dc.date.available | 2019-05-20T10:00:37Z | |
dc.date.issued | 2019-05-08 | |
dc.identifier.citation | Mol Ther. 2019 ;27(5):986-998. | es_ES |
dc.identifier.issn | 15250016 | es_ES |
dc.identifier.uri | http://hdl.handle.net/20.500.12105/7611 | |
dc.description.abstract | Gene editing constitutes a novel approach for precisely correcting disease-causing gene mutations. Frameshift mutations in COL7A1 causing recessive dystrophic epidermolysis bullosa are amenable to open reading frame restoration by non-homologous end joining repair-based approaches. Efficient targeted deletion of faulty COL7A1 exons in polyclonal patient keratinocytes would enable the translation of this therapeutic strategy to the clinic. In this study, using a dual single-guide RNA (sgRNA)-guided Cas9 nuclease delivered as a ribonucleoprotein complex through electroporation, we have achieved very efficient targeted deletion of COL7A1 exon 80 in recessive dystrophic epidermolysis bullosa (RDEB) patient keratinocytes carrying a highly prevalent frameshift mutation. This ex vivo non-viral approach rendered a large proportion of corrected cells producing a functional collagen VII variant. The effective targeting of the epidermal stem cell population enabled long-term regeneration of a properly adhesive skin upon grafting onto immunodeficient mice. A safety assessment by next-generation sequencing (NGS) analysis of potential off-target sites did not reveal any unintended nuclease activity. Our strategy could potentially be extended to a large number of COL7A1 mutation-bearing exons within the long collagenous domain of this gene, opening the way to precision medicine for RDEB. | es_ES |
dc.description.sponsorship | The study was mainly supported by DEBRA International, funded byDEBRA Austria (grant termed as Larcher 1). Additional funds camefrom Spanish grants SAF2017-86810-R (to M.D.R.) and PI17/01747(to F.L.) from the Ministry of Economy and Competitiveness and In-stituto de Salud Carlos III, respectively, both co-funded with Euro-pean Regional Development Funds (ERDF) ERA-NET E-RAREJTC 2017 (MutaEB) and CIBERER (grant termed as Murillas-TERAPIAS ER2017). Authors are indebted to Blanca Duarte, Almu-dena Holguín, and Nuria Illera for grafting experiments and to JesusMartínez and Edilia De Almeida for animal maintenance and care.We also thank Juan Manuel Ruibal Mera for the suction blister devicemanufacturing and Jonathan O’Keeffe for professional language ex-amination of the manuscript | es_ES |
dc.language.iso | eng | es_ES |
dc.publisher | Elsevier | es_ES |
dc.type.hasVersion | VoR | es_ES |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-sa/4.0/ | * |
dc.subject | CRISPR/Cas9 | es_ES |
dc.subject | Epidermal stem cells | es_ES |
dc.subject | Epidermolysis bullosa | es_ES |
dc.subject | Gene therapy | es_ES |
dc.title | Clinically Relevant Correction of Recessive Dystrophic Epidermolysis Bullosa by Dual sgRNA CRISPR/Cas9-Mediated Gene Editing | es_ES |
dc.type | journal article | es_ES |
dc.rights.license | Atribución-NoComercial-CompartirIgual 4.0 Internacional | * |
dc.identifier.pubmedID | 30930113 | es_ES |
dc.format.volume | 27 | es_ES |
dc.format.number | 5 | es_ES |
dc.format.page | 986-998 | es_ES |
dc.identifier.doi | 10.1016/j.ymthe.2019.03.007 | es_ES |
dc.contributor.funder | Ministerio de Economía y Competitividad (España) | |
dc.contributor.funder | Instituto de Salud Carlos III | |
dc.contributor.funder | DEBRA Austria | |
dc.description.peerreviewed | Sí | es_ES |
dc.identifier.e-issn | 1525-0024 | es_ES |
dc.relation.publisherversion | https://doi.org/10.1016/j.ymthe.2019.03.007. | es_ES |
dc.identifier.journal | Molecular therapy : the journal of the American Society of Gene Therapy | es_ES |
dc.repisalud.institucion | CNIO | es_ES |
dc.repisalud.orgCNIO | CNIO::Unidades técnicas::Unidad de Citogenética Molecular | es_ES |
dc.relation.projectID | info:eu-repo/grantAgreement/ES/AF2017-86810-R | es_ES |
dc.relation.projectID | info:eu-repo/grantAgreement/ES/PI17/01747 | es_ES |
dc.rights.accessRights | open access | es_ES |