title: Recombinant adeno-associated viral vectors.
abstract: Recombinant adeno-associated viral (rAAV) vectors can be used to locally or systemically enhance or silence gene expression. They are relatively nonimmunogenic and can transduce dividing and nondividing cells, and different rAAV serotypes may transduce diverse cell types. Therefore, rAAV vectors are excellent tools to study the function of neuropeptides in local brain areas. In this chapter, we describe a protocol to produce high-titer, in vivo grade, rAAV vector stocks. The protocol includes an Iodixanol gradient, an anion exchange column and a desalting/concentration step and can be used for every serotype. In addition, a short protocol for rAAV injections into the brain and directions on how to detect and localize transduced cells are given.
authors.affiliation: Department of Neuroscience and Pharmacology, Rudolf Magnus Institute of Neuroscience, Utrecht University Medical Centre Utrecht, Utrecht, The Netherlands.
authors.forename: Marijke W A
authors.initials: MW
authors.lastname: de Backer
authors.forename: Keith M
authors.initials: KM
authors.lastname: Garner
authors.forename: Mieneke C M
authors.initials: MC
authors.lastname: Luijendijk
authors.forename: Roger A H
authors.initials: RA
authors.lastname: Adan
mesh_terms: D002460:Cell Line; D002490:Central Nervous System / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction; D014283:Triiodobenzoic Acids / Q000737:chemistry
chemical_list: D014283:Triiodobenzoic Acids; C044834:iodixanol