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Gene editing for the efficient correction of a recurrent COL7A1 mutation in recessive dystrophic epidermolysis bullosa keratinocytes

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dc.affiliation.dptoUC3M. Departamento de Bioingenieríaes
dc.affiliation.grupoinvUC3M. Grupo de Investigación: Tissue Engineering and Regenerative Medicine (TERMeG)es
dc.contributor.authorChamorro, Cristina
dc.contributor.authorMencía Rodríguez, Ángeles
dc.contributor.authorAlmarza, David
dc.contributor.authorDuarte, Blanca
dc.contributor.authorBüning, Hildegard
dc.contributor.authorSallach, Jessica
dc.contributor.authorHausser, Ingrid
dc.contributor.authorRío Nechaevsky, Marcela del
dc.contributor.authorLarcher Laguzzi, Fernando
dc.contributor.authorMurillas, Rodolfo
dc.contributor.funderComunidad de Madrides
dc.contributor.funderMinisterio de Economía y Competitividad (España)es
dc.date.accessioned2023-11-21T11:52:59Z
dc.date.available2023-11-21T11:52:59Z
dc.date.issued2016-01
dc.description.abstractClonal gene therapy protocols based on the precise manipulation of epidermal stem cells require highly efficient gene-editing molecular tools. We have combined adeno-associated virus (AAV)-mediated delivery of donor template DNA with transcription activator-like nucleases (TALE) expressed by adenoviral vectors to address the correction of the c.6527insC mutation in the COL7A1 gene, causing recessive dystrophic epidermolysis bullosa in a high percentage of Spanish patients. After transduction with these viral vectors, high frequencies of homology-directed repair were found in clones of keratinocytes derived from a recessive dystrophic epidermolysis bullosa (RDEB) patient homozygous for the c.6527insC mutation. Gene-edited clones recovered the expression of the COL7A1 transcript and collagen VII protein at physiological levels. In addition, treatment of patient keratinocytes with TALE nucleases in the absence of a donor template DNA resulted in nonhomologous end joining (NHEJ)-mediated indel generation in the vicinity of the c.6527insC mutation site in a large proportion of keratinocyte clones. A subset of these indels restored the reading frame of COL7A1 and resulted in abundant, supraphysiological expression levels of mutant or truncated collagen VII protein. Keratinocyte clones corrected both by homology-directed repair (HDR) or NHEJ were used to regenerate skin displaying collagen VII in the dermo-epidermal junction.en
dc.description.sponsorshipThis work was supported in part by grants SAF2014-54885-R from MINECO to MDR; S2010/BMD-2420 and S2010/ BMD-2359 from Dirección General de Investigación de la Comunidad de Madrid to MDR and FL respectively; PI14/00931 from Instituto de Salud Carlos III to FL and by funding from Center for Molecular Medicine Cologne (CMMC) of the University of Cologne to H.B. There are no conflicts of interest.en
dc.format.extent13
dc.identifier.bibliographicCitationChamorro, C., Mencía, Á., Almarza, D., Duarte, B., Büning, H., Sallach, J., Hausser, I., Del Río, M., Larcher, F., & Murillas, R. (2016). Gene editing for the efficient correction of a recurrent COL7A1 mutation in recessive dystrophic epidermolysis bullosa keratinocytes. Molecular Therapy - Nucleic Acids, 5, e307.en
dc.identifier.doihttps://doi.org/10.1038/mtna.2016.19
dc.identifier.issn2162-2531
dc.identifier.publicationfirstpage1
dc.identifier.publicationissuee307
dc.identifier.publicationlastpage13
dc.identifier.publicationtitleMolecular Therapy-Nucleic Acidsen
dc.identifier.publicationvolume5
dc.identifier.urihttps://hdl.handle.net/10016/38921
dc.identifier.uxxiAR/0000019340
dc.language.isoengen
dc.publisherElsevieren
dc.relation.projectIDComunidad de Madrid. S2010/ BMD-2359es
dc.relation.projectIDComunidad de Madrid. S2010/BMD-2420es
dc.relation.projectIDGobierno de España. SAF2014-54885-Res
dc.rights© 2016 Official journal of the American Society of Gene & Cell Therapy. Published by Elsevier Inc.en
dc.rightsAtribución-NoComercial-CompartirIgual 3.0 España*
dc.rights.accessRightsopen accessen
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/3.0/es/*
dc.subject.ecienciaBiología y Biomedicinaes
dc.subject.ecienciaInformáticaes
dc.subject.ecienciaMedicinaes
dc.subject.otherAAV vectorsen
dc.subject.otherEpidermolysis bullosaen
dc.subject.otherGene editingen
dc.subject.otherIndelsen
dc.subject.otherHomologous recombinationen
dc.subject.otherTalenen
dc.titleGene editing for the efficient correction of a recurrent COL7A1 mutation in recessive dystrophic epidermolysis bullosa keratinocytesen
dc.typeresearch article*
dc.type.hasVersionVoR*
dspace.entity.typePublication
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