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dc.contributor.authorKrooss, Simon Alexander
dc.contributor.authorDai, Zhen
dc.contributor.authorSchmidt, Florian
dc.contributor.authorRovai, Alice
dc.contributor.authorFakhiri, Julia
dc.contributor.authorDhingra, Akshay
dc.contributor.authorYuan, Qinggong
dc.contributor.authorYang, Taihua
dc.contributor.authorBalakrishnan, Asha
dc.contributor.authorSteinbrück, Lars
dc.contributor.authorSrivaratharajan, Sangar
dc.contributor.authorManns, Michael Peter
dc.contributor.authorSchambach, Axel
dc.contributor.authorGrimm, Dirk
dc.contributor.authorBohne, Jens
dc.contributor.authorSharma, Amar Deep
dc.contributor.authorBüning, Hildegard
dc.contributor.authorOtt, Michael
dc.date.accessioned2020-01-31T09:29:54Z
dc.date.available2020-01-31T09:29:54Z
dc.date.issued2020-01-24
dc.identifier.citationiScience. 2020 Jan 24;23(1):100764. doi: 10.1016/j.isci.2019.100764. Epub 2019 Dec 12.en_US
dc.identifier.issn2589-0042
dc.identifier.pmid31887661
dc.identifier.doi10.1016/j.isci.2019.100764
dc.identifier.urihttp://hdl.handle.net/10033/622113
dc.description.abstractAdeno-associated virus (AAV)-based vectors are considered efficient and safe gene delivery systems in gene therapy. We combined two guide RNA genes, Cas9, and a self-linearizing repair template in one vector (AIO-SL) to correct fumarylacetoacetate hydrolase (FAH) deficiency in mice. The vector genome of 5.73 kb was packaged into VP2-depleted AAV particles (AAV2/8ΔVP2), which, however, did not improve cargo capacity. Reprogrammed hepatocytes were treated with AIO-SL.AAV2ΔVP2 and subsequently transplanted, resulting in large clusters of FAH-positive hepatocytes. Direct injection of AIO-SL.AAV8ΔVP2 likewise led to FAH expression and long-term survival. The AIO-SL vector achieved an ∼6-fold higher degree of template integration than vectors without template self-linearization. Subsequent analysis revealed that AAV8 particles, in contrast to AAV2, incorporate oversized genomes distinctly greater than 5.2 kb. Finally, our AAV8-based vector represents a promising tool for gene editing strategies to correct monogenic liver diseases requiring (large) fragment removal and/or simultaneous sequence replacement.en_US
dc.language.isoenen_US
dc.publisherCell Press/Elsevieren_US
dc.rightsAttribution-NonCommercial-ShareAlike 4.0 International*
dc.rights.urihttp://creativecommons.org/licenses/by-nc-sa/4.0/*
dc.subjectGenetic Engineeringen_US
dc.subjectGeneticsen_US
dc.subjectTechniques in Geneticsen_US
dc.titleEx Vivo/In vivo Gene Editing in Hepatocytes Using "All-in-One" CRISPR-Adeno-Associated Virus Vectors with a Self-Linearizing Repair Template.en_US
dc.typeArticleen_US
dc.contributor.departmentTWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany.en_US
dc.identifier.journaliScienceen_US
refterms.dateFOA2020-01-31T09:29:55Z
dc.source.journaltitleiScience


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