A Self-Deleting AAV-CRISPR System for In Vivo Genome Editing

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dc.citation.firstpage111en_US
dc.citation.journalTitleMolecular Therapy - Methods & Clinical Developmenten_US
dc.citation.lastpage122en_US
dc.citation.volumeNumber12en_US
dc.contributor.authorLi, Angen_US
dc.contributor.authorLee, Ciaran M.en_US
dc.contributor.authorHurley, Ayrea E.en_US
dc.contributor.authorJarrett, Kelsey E.en_US
dc.contributor.authorDe Giorgi, Marcoen_US
dc.contributor.authorLu, Weiqien_US
dc.contributor.authorBalderrama, Karol S.en_US
dc.contributor.authorDoerfler, Alexandria M.en_US
dc.contributor.authorDeshmukh, Harshavardhanen_US
dc.contributor.authorRay, Anirbanen_US
dc.contributor.authorBao, Gangen_US
dc.contributor.authorLagor, William R.en_US
dc.date.accessioned2019-01-24T16:07:54Zen_US
dc.date.available2019-01-24T16:07:54Zen_US
dc.date.issued2019en_US
dc.description.abstractAdeno-associated viral (AAV) vectors packaging the CRISPR-Cas9 system (AAV-CRISPR) can efficiently modify disease-relevant genes in somatic tissues with high efficiency. AAV vectors are a preferred delivery vehicle for tissue-directed gene therapy because of their ability to achieve sustained expression from largely non-integrating episomal genomes. However, for genome editizng applications, permanent expression of non-human proteins such as the bacterially derived Cas9 nuclease is undesirable. Methods are needed to achieve efficient genome editing in vivo, with controlled transient expression of CRISPR-Cas9. Here, we report a self-deleting AAV-CRISPR system that introduces insertion and deletion mutations into AAV episomes. We demonstrate that this system dramatically reduces the level of Staphylococcus aureus Cas9 protein, often greater than 79%, while achieving high rates of on-target editing in the liver. Off-target mutagenesis was not observed for the self-deleting Cas9 guide RNA at any of the predicted potential off-target sites examined. This system is efficient and versatile, as demonstrated by robust knockdown of liver-expressed proteins in vivo. This self-deleting AAV-CRISPR system is an important proof of concept that will help enable translation of liver-directed genome editing in humans.en_US
dc.identifier.citationLi, Ang, Lee, Ciaran M., Hurley, Ayrea E., et al.. "A Self-Deleting AAV-CRISPR System for In Vivo Genome Editing." <i>Molecular Therapy - Methods & Clinical Development,</i> 12, (2019) Elsevier: 111-122. https://doi.org/10.1016/j.omtm.2018.11.009.en_US
dc.identifier.digitalAAV-CRISPRen_US
dc.identifier.doihttps://doi.org/10.1016/j.omtm.2018.11.009en_US
dc.identifier.urihttps://hdl.handle.net/1911/105116en_US
dc.language.isoengen_US
dc.publisherElsevieren_US
dc.rightsThis is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).en_US
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/en_US
dc.subject.keywordAAVen_US
dc.subject.keywordAAV-CRISPRen_US
dc.subject.keywordCRISPR/Cas9en_US
dc.subject.keywordadeno-associated virusen_US
dc.subject.keywordgene therapyen_US
dc.subject.keywordin vivo deliveryen_US
dc.subject.keywordliveren_US
dc.subject.keywordself-deletingen_US
dc.subject.keywordsomatic genome editingen_US
dc.titleA Self-Deleting AAV-CRISPR System for In Vivo Genome Editingen_US
dc.typeJournal articleen_US
dc.type.dcmiTexten_US
dc.type.publicationpublisher versionen_US
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