Random Insertion of mCherry Into VP3 Domain of Adeno- associated Virus Yields Fluorescent Capsids With no Loss of Infectivity

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dc.citation.firstpagee54
dc.citation.journalTitleMolecular TherapyﾖNucleic Acids
dc.citation.volumeNumber1
dc.contributor.authorJudd, Justin
dc.contributor.authorWei, Fang
dc.contributor.authorNguyen, Peter Q.
dc.contributor.authorTartaglia, Lawrence J.
dc.contributor.authorAgbandje-McKenna, Mavis
dc.contributor.authorSilberg, Jonathan J.
dc.contributor.authorSuh, Junghae
dc.date.accessioned2013-01-29T20:21:32Z
dc.date.available2013-01-29T20:21:32Z
dc.date.issued2012
dc.description.abstractAdeno-associated virus (AAV)-derived vectors are promising gene delivery systems, and a number of design strategies have been pursued to improve their performance. For example, genetic insertion of proteins into the capsid may be used to achieve vector retargeting, reduced immunogenicity, or to track vector transport. Unfortunately, rational approaches to genetic insertion have experienced limited success due to the unpredictable context-dependent nature of protein folding and the complexity of the capsid's macroassembly. We report the construction and use of a frame-enriched DNase-based random insertion library based on AAV2 cap, called pAAV2_RaPID (Random Peptide Insertion by DNase). The fluorescent mCherry protein was inserted randomly throughout the AAV2 capsid and the library was selected for fluorescent and infectious variants. A capsid site was identified in VP3 that can tolerate the large protein insertion. In contrast to previous efforts to incorporate fluorescent proteins into the AAV2 capsid, the isolated mCherry mutant maintains native infectivity while displaying robust fluorescence. Collectively, these results demonstrate that the pAAV2_RaPID platform library can be used to create fully infectious AAV vectors carrying large functional protein domains on the capsid.
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dc.identifier.citationJudd, Justin, Wei, Fang, Nguyen, Peter Q., et al.. "Random Insertion of mCherry Into VP3 Domain of Adeno- associated Virus Yields Fluorescent Capsids With no Loss of Infectivity." <i>Molecular TherapyﾖNucleic Acids,</i> 1, (2012) American Society of Gene & Cell Therapy: e54. http://dx.doi.org/10.1038/mtna.2012.46.
dc.identifier.doihttp://dx.doi.org/10.1038/mtna.2012.46
dc.identifier.urihttps://hdl.handle.net/1911/69863
dc.language.isoeng
dc.publisherAmerican Society of Gene & Cell Therapy
dc.rightsThis work is licensed under the Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 Unported License.
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/3.0/
dc.titleRandom Insertion of mCherry Into VP3 Domain of Adeno- associated Virus Yields Fluorescent Capsids With no Loss of Infectivity
dc.typeJournal article
dc.type.dcmiText
dc.type.publicationpublisher version
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