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

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dc.citation.firstpage111
dc.citation.journalTitleMolecular Therapy - Methods & Clinical Development
dc.citation.lastpage122
dc.citation.volumeNumber12
dc.contributor.authorLi, Ang
dc.contributor.authorLee, Ciaran M.
dc.contributor.authorHurley, Ayrea E.
dc.contributor.authorJarrett, Kelsey E.
dc.contributor.authorDe Giorgi, Marco
dc.contributor.authorLu, Weiqi
dc.contributor.authorBalderrama, Karol S.
dc.contributor.authorDoerfler, Alexandria M.
dc.contributor.authorDeshmukh, Harshavardhan
dc.contributor.authorRay, Anirban
dc.contributor.authorBao, Gang
dc.contributor.authorLagor, William R.
dc.date.accessioned2019-01-24T16:07:54Z
dc.date.available2019-01-24T16:07:54Z
dc.date.issued2019
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.
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.
dc.identifier.digitalAAV-CRISPR
dc.identifier.doihttps://doi.org/10.1016/j.omtm.2018.11.009
dc.identifier.urihttps://hdl.handle.net/1911/105116
dc.language.isoeng
dc.publisherElsevier
dc.rightsThis is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject.keywordAAV
dc.subject.keywordAAV-CRISPR
dc.subject.keywordCRISPR/Cas9
dc.subject.keywordadeno-associated virus
dc.subject.keywordgene therapy
dc.subject.keywordin vivo delivery
dc.subject.keywordliver
dc.subject.keywordself-deleting
dc.subject.keywordsomatic genome editing
dc.titleA Self-Deleting AAV-CRISPR System for In Vivo Genome Editing
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpublisher version
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