Ex Vivo/In vivo Gene Editing in Hepatocytes Using "All-in-One" CRISPR-Adeno-Associated Virus Vectors with a Self-Linearizing Repair Template.
dc.contributor.author | Krooss, Simon Alexander | |
dc.contributor.author | Dai, Zhen | |
dc.contributor.author | Schmidt, Florian | |
dc.contributor.author | Rovai, Alice | |
dc.contributor.author | Fakhiri, Julia | |
dc.contributor.author | Dhingra, Akshay | |
dc.contributor.author | Yuan, Qinggong | |
dc.contributor.author | Yang, Taihua | |
dc.contributor.author | Balakrishnan, Asha | |
dc.contributor.author | Steinbrück, Lars | |
dc.contributor.author | Srivaratharajan, Sangar | |
dc.contributor.author | Manns, Michael Peter | |
dc.contributor.author | Schambach, Axel | |
dc.contributor.author | Grimm, Dirk | |
dc.contributor.author | Bohne, Jens | |
dc.contributor.author | Sharma, Amar Deep | |
dc.contributor.author | Büning, Hildegard | |
dc.contributor.author | Ott, Michael | |
dc.date.accessioned | 2020-01-31T09:29:54Z | |
dc.date.available | 2020-01-31T09:29:54Z | |
dc.date.issued | 2020-01-24 | |
dc.identifier.citation | iScience. 2020 Jan 24;23(1):100764. doi: 10.1016/j.isci.2019.100764. Epub 2019 Dec 12. | en_US |
dc.identifier.issn | 2589-0042 | |
dc.identifier.pmid | 31887661 | |
dc.identifier.doi | 10.1016/j.isci.2019.100764 | |
dc.identifier.uri | http://hdl.handle.net/10033/622113 | |
dc.description.abstract | Adeno-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.iso | en | en_US |
dc.publisher | Cell Press/Elsevier | en_US |
dc.rights | Attribution-NonCommercial-ShareAlike 4.0 International | * |
dc.rights.uri | http://creativecommons.org/licenses/by-nc-sa/4.0/ | * |
dc.subject | Genetic Engineering | en_US |
dc.subject | Genetics | en_US |
dc.subject | Techniques in Genetics | en_US |
dc.title | Ex 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.type | Article | en_US |
dc.contributor.department | TWINCORE, Zentrum für experimentelle und klinische Infektionsforschung GmbH,Feodor-Lynen Str. 7, 30625 Hannover, Germany. | en_US |
dc.identifier.journal | iScience | en_US |
refterms.dateFOA | 2020-01-31T09:29:55Z | |
dc.source.journaltitle | iScience |