AAV2 vectors and methods

DETAILED DESCRIPTION The present invention is illustrated by the following examples that are not intended to limit the invention.
Example I describes construction of AAV packaging plasmids encoding altered capsid proteins and analysis of the ability of the altered capsid proteins to be assembled into infectious AAV vectors.
Example 2 presents assays for the surface expression of epitopes inserted in the altered capsid proteins.
Example 3 describes experiments testing whether the AAV vectors retained HSPG-binding ability.
Example 4 describes construction and characterization of a mutant AAV vector containing a double insertion within the capsid protein.
Example 5 includes analysis of the effect of linker and scaffold sequences on the altered capsid proteins.
Example 6 presents the results of experiments in which AAV vectors encoding capsid proteins with an insertion of an luteinizing hormone receptor binding peptide were able to transduce OVCAR-3 cells.
Example 6 also discusses various indications amenable to use of AAV vectors of the invention.
Example 7 and 8 describe fourteen additional modified AAV vectors, wherein the RGD-4C peptide motif was inserted into the capsid proteins.
The experiments described in Example 9 demonstrate that the AAV-RGD vectors attach to and enter cells via integrin receptors.
Example 10 demonstrates that the AAV-RGD vectors were capable of mediating gene delivery via integrin receptors.
Example demonstrates that the AAV-RGD vectors transferred genes to ovarian adenocarcinoma cell lines.
Example 12 describes AAV mediated eGFP gene delivery to human ovarian tumor xenografts established in SCID mice.
Example 13 describes construction of mutant AAV vectors which are biotinylated in vivo through an insertion of the biotin acceptor peptide in the capsid protein.
Finally, Example 14 describes a packaging system for biotinylated AAV vectors.
EXAMPLE 1 In order to identify sites within the AAV2 capsid that could tolerate insertion of targeting epitopes, an extensive site-specific mutagenesis strategy was designed