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Vector Systems for Prenatal Gene Therapy: Principles of Adeno-Associated Virus Vector Design and Production

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Prenatal Gene Therapy

Part of the book series: Methods in Molecular Biology ((MIMB,volume 891))

Abstract

Vectors based on adeno-associated virus (AAV) show great promise for safe, efficacious therapeutic gene transfer in extensive pre-clinical data and, recently, in clinical trials. Careful vector design and choice from a range of natural or synthetic pseudotypes allow targeted, efficient, and sustained expression of therapeutic genes. The efficiency of gene delivery can be further enhanced through the use of drug pre-treatment or co-infection with a suitable helper virus. This chapter describes current best practice for AAV production, including complete methods for: (1) efficient generation of vector without the use of helper viruses, simplifying the transition to GMP-grade production for clinical applications; (2) efficient and easily scalable purification of the virus by affinity chromatography, allowing rapid production of highly concentrated, high titre stocks; (3) reliable quantification and assaying of viral stocks, along with short- and long-term storage considerations.

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Author information

Authors and Affiliations

  1. Department of Haematology, UCL Cancer Institute, London, UK

    Christopher J. Binny & Amit C. Nathwani MD, PhD

Authors
  1. Christopher J. Binny
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  2. Amit C. Nathwani MD, PhD
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Corresponding author

Correspondence to Amit C. Nathwani MD, PhD .

Editor information

Editors and Affiliations

  1. National Heart & Lung Inst., Molecular & Cellular Medicine Sec., Imperial College London, Imperial College Road, London, SW7 2AZ, United Kingdom

    Charles Coutelle

  2. Reader in Gene Transfer Technology, Gene Transfer Technology Group, University College London, Chenies Mews 86-96, London, WC1E 6HX, United Kingdom

    Simon N. Waddington

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Binny, C.J., Nathwani, A.C. (2012). Vector Systems for Prenatal Gene Therapy: Principles of Adeno-Associated Virus Vector Design and Production. In: Coutelle, C., Waddington, S. (eds) Prenatal Gene Therapy. Methods in Molecular Biology, vol 891. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-873-3_6

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  • DOI: https://doi.org/10.1007/978-1-61779-873-3_6

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-61779-872-6

  • Online ISBN: 978-1-61779-873-3

  • eBook Packages: Springer Protocols

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