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Recombinant AAV Delivery to the Central Nervous System

  • Olivier Bockstael
  • Kevin D. Foust
  • Brian Kaspar
  • Liliane TenenbaumEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 807)

Abstract

Recombinant AAV-mediated gene delivery to the CNS can be performed either by direct delivery at the target site or from the periphery, using intramuscular injections and retrograde transport along motor neuron projections or intravenous injections and blood–brain barrier crossing.

In this chapter, we describe:
  1. 1.

    Methods for recombinant virus administration, including stereotactic surgery, intramuscular, and intravenous administration.

     
  2. 2.

    Methods to evaluate the number and biodistribution of brain and spinal cord cells expressing the transgene by immunohistochemisty as well as the amount of transgene product by ELISA in the target region.

     
  3. 3.

    Methods to characterize the cellular specificity of transgene expression by double immunofluorescence.

     
  4. 4.

    Methods to quantify the amounts of viral DNA as well as of transgene mRNA by quantitative PCR and RT-PCR, respectively.

     

Key words

Stereotaxy Intravenous injection Retrograde transport Immunohistochemistry Confocal microscopy GFP GDNF Quantitative PCR mRNA Hirt DNA 

Notes

Acknowledgments

O.B. was the recipient of a predoctoral fellowship from the Belgian “FRIA” (Fonds pour la Recherche dans l’Industrie et l’Agriculture and FNRS-Télévie). L.T. was the recipient of a “Crédit aux chercheurs” from the Belgian National Research Foundation. This work was also supported by grants from “Fonds National de la Recherche Scientifique Médicale,” “Région Bruxelles-Capitale,” and “Association Française contre les Myopathies.”

The Kaspar Laboratory is funded in part from The National Institute of Health (NIH).

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Copyright information

© Springer Science+Business Media, LLC 2012

Authors and Affiliations

  • Olivier Bockstael
    • 1
  • Kevin D. Foust
    • 2
  • Brian Kaspar
    • 2
  • Liliane Tenenbaum
    • 1
    • 3
    Email author
  1. 1.Hôpital ErasmeUniversité Libre de BruxellesBrusselsBelgium
  2. 2.The Research Institute at Nationwide Children’s HospitalColumbusUSA
  3. 3.Lausanne Hospital UniversityLausanneSwitzerland

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