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Design and Construction of Functional AAV Vectors

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Adeno-Associated Virus

Part of the book series: Methods in Molecular Biology ((MIMB,volume 807))

Abstract

Using the basic principles of molecular biology and laboratory techniques presented in this chapter, researchers should be able to create a wide variety of AAV vectors for both clinical and basic research applications. Basic vector design concepts are covered for both protein coding gene expression and small non-coding RNA gene expression cassettes. AAV plasmid vector backbones (available via AddGene) are described, along with critical sequence details for a variety of modular expression components that can be inserted as needed for specific applications. Protocols are provided for assembling the various DNA components into AAV vector plasmids in Escherichia coli, as well as for transferring these vector sequences into baculovirus genomes for large-scale production of AAV in the insect cell production system.

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Author information

Authors and Affiliations

  1. Department of Hematology, St. Jude Children’s Research Hospital, Memphis, TN, USA

    John T. Gray

  2. Division of Cellular and Molecular Therapy, University of Florida, Gainesville, FL, USA

    Serge Zolotukhin

Authors
  1. John T. Gray
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  2. Serge Zolotukhin
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Correspondence to John T. Gray .

Editor information

Editors and Affiliations

  1. Center of Excellence for Regenerative, Health Biotechnology (CERHB), University of Florida, Innovation Drive 13706, Alachua, 32615, Florida, USA

    Richard O. Snyder

  2. INSERM U649, Labo. Thérapie Génique, CHU Hôtel Dieu, bd. Jean Monnet 30, Nantes CX 1, 44035, France

    Philippe Moullier

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Gray, J.T., Zolotukhin, S. (2012). Design and Construction of Functional AAV Vectors. In: Snyder, R., Moullier, P. (eds) Adeno-Associated Virus. Methods in Molecular Biology, vol 807. Humana Press. https://doi.org/10.1007/978-1-61779-370-7_2

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  • DOI: https://doi.org/10.1007/978-1-61779-370-7_2

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-369-1

  • Online ISBN: 978-1-61779-370-7

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