Recombinant AAV delivery to the central nervous system.

Details

Serval ID
serval:BIB_38DBA13884EB
Type
A part of a book
Publication sub-type
Chapter: chapter ou part
Collection
Publications
Institution
Title
Recombinant AAV delivery to the central nervous system.
Title of the book
Methods Mol Biol
Author(s)
Bockstael O., Foust K.D., Kaspar B., Tenenbaum L.
Publisher
Human Press
ISBN
1940-6029 (Electronic)
Publication state
Published
Issued date
2011
Volume
807
Pages
159-177
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't Publication Status: ppublish
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. Methods for recombinant virus administration, including stereotactic surgery, intramuscular, and intravenous administration. 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. Methods to characterize the cellular specificity of transgene expression by double immunofluorescence. 4. Methods to quantify the amounts of viral DNA as well as of transgene mRNA by quantitative PCR and RT-PCR, respectively.
Keywords
Animals, Central Nervous System/metabolism, Dependovirus/genetics, Genetic Vectors/genetics, Immunohistochemistry, Mice, Microscopy, Confocal, Motor Neurons/metabolism, Polymerase Chain Reaction
Pubmed
Create date
07/02/2013 14:45
Last modification date
21/08/2019 5:35
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