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:
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1.
Methods for recombinant virus administration, including stereotactic surgery, intramuscular, and intravenous administration.
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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.
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3.
Methods to characterize the cellular specificity of transgene expression by double immunofluorescence.
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4.
Methods to quantify the amounts of viral DNA as well as of transgene mRNA by quantitative PCR and RT-PCR, respectively.
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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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Bockstael, O., Foust, K.D., Kaspar, B., Tenenbaum, L. (2012). Recombinant AAV Delivery to the Central Nervous System. 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_7
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DOI: https://doi.org/10.1007/978-1-61779-370-7_7
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