In vitro selection of viral vectors with modified tropism: the adeno-associated virus display.
Improving the efficiency and specificity of gene vectors is critical for the success of gene therapy. In an effort to generate viral mutants with controlled tropism we produced a library of adeno-associated virus (AAV) clones with randomly modified capsids and used it for the selection of receptor-targeting mutants. After several rounds of selection on different cell lines that were resistant to infection by wild-type (wt) AAV, infectious mutants were harvested at high titers. These mutants transduced target cells with an up to 100-fold increased efficiency, in a receptor-specific manner and without interacting with the primary receptor for wt AAV. The results demonstrate for the first time that a combinatorial approach based on a eukaryotic virus library allows one to generate efficient, receptor-specific targeting vectors with desired tropism.
Perabo
Luca
L
Genzentrum, Ludwig-Maximilians-Universität München, Feodor-Lynen-Strasse 25, 81377, Munich, Germany.
Büning
Hildegard
H
Kofler
David M
DM
Ried
Martin U
MU
Girod
Anne
A
Wendtner
Clemens M
CM
Enssle
Jörg
J
Hallek
Michael
M
D001483:Base Sequence; D002213:Capsid / Q000378:metabolism; D002460:Cell Line; D002469:Cell Separation; D000229:Dependovirus / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D015723:Gene Library; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000032:analysis*; D006367:HeLa Cells; D006801:Humans; D066298:In Vitro Techniques; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000378:metabolism; D008954:Models, Biological; D008969:Molecular Sequence Data; D009154:Mutation; D010957:Plasmids / Q000378:metabolism; D014161:Transduction, Genetic; D018523:Tropism