A survey of ex vivo/in vitro transduction efficiency of mammalian primary cells and cell lines with Nine natural adeno-associated virus (AAV1-9) and one engineered adeno-associated virus serotype Public Deposited

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Creator
  • Ellis, Brian L
    • Other Affiliation: Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX, USA
  • Steininger, Robert J
    • Other Affiliation: Department of Pharmacology, Green Center for Systems Biology, Simmons Cancer Center, University of Texas Southwestern Medical Center, Dallas, TX, USA
  • Connelly, Jon P
    • Other Affiliation: Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX, USA
  • Hirsch, Matthew L
    • Affiliation: School of Medicine, Department of Ophthalmology, Gene Therapy Center
  • Barker, Jenny C
    • Other Affiliation: Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX, USA
  • Porteus, Matthew H
    • Other Affiliation: Department of Biochemistry, University of Texas Southwestern Medical Center, Dallas, TX, USA; Department of Pediatrics, University of Texas Southwestern Medical Center, Dallas, TX, 75390-9148, USA
Abstract
  • Abstract: Background: The ability to deliver a gene of interest into a specific cell type is an essential aspect of biomedical research. Viruses can be a useful tool for this delivery, particularly in difficult to transfect cell types. Adeno-associated virus (AAV) is a useful gene transfer vector because of its ability to mediate efficient gene transduction in numerous dividing and quiescent cell types, without inducing any known pathogenicity. There are now a number of natural for that designed AAV serotypes that each has a differential ability to infect a variety of cell types. Although transduction studies have been completed, the bulk of the studies have been done in vivo, and there has never been a comprehensive study of transduction ex vivo/in vitro. Methods: Each cell type was infected with each serotype at a multiplicity of infection of 100,000 viral genomes/cell and transduction was analyzed by flow cytometry + . Results: We found that AAV1 and AAV6 have the greatest ability to transduce a wide range of cell types, however, for particular cell types, there are specific serotypes that provide optimal transduction. Conclusions: In this work, we describe the transduction efficiency of ten different AAV serotypes in thirty-four different mammalian cell lines and primary cell types. Although these results may not be universal due to numerous factors such as, culture conditions and/ or cell growth rates and cell heterogeneity, these results provide an important and unique resource for investigators who use AAV as an ex vivo gene delivery vector or who work with cells that are difficult to transfect.
Date of publication
Identifier
  • doi:10.1186/1743-422X-10-74
  • 23497173
Resource type
  • Article
Rights statement
  • In Copyright
Rights holder
  • Brian L Ellis et al.; licensee BioMed Central Ltd.
License
Journal title
  • Virology Journal
Journal volume
  • 10
Journal issue
  • 1
Page start
  • 74
Language
  • English
Is the article or chapter peer-reviewed?
  • Yes
ISSN
  • 1743-422X
Bibliographic citation
  • Virology Journal. 2013 Mar 06;10(1):74
Access
  • Open Access
Publisher
  • BioMed Central Ltd
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