Human gene targeting by viral vectors.
Stable transduction of mammalian cells typically involves random integration of viral vectors by non-homologous recombination. Here we report that vectors based on adeno-associated virus (AAV) can efficiently modify homologous human chromosomal target sequences. Both integrated neomycin phosphotransferase genes and the hypoxanthine phosphoribosyltransferase gene were targeted by AAV vectors. Site-specific genetic modifications could be introduced into approximately 1% of cells, with the highest targeting rates occurring in normal human fibroblasts. These results suggest that AAV vectors could be used to introduce specific genetic changes into the genomic DNA of a wide variety of mammalian cells, including therapeutic gene targeting applications.
Russell
D W
DW
Markey Molecular Medicine Center, Department of Medicine, University of Washington School of Medicine, Seattle 98195-7720, USA. drussell@u.washington.edu
Hirata
R K
RK
D000256:Adenoviridae / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D018390:Gene Targeting; D018014:Gene Transfer Techniques; D005796:Genes / Q000235:genetics; D005814:Genes, Viral / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics* / Q000494:pharmacology; D006801:Humans; D007041:Hypoxanthine Phosphoribosyltransferase / Q000235:genetics; D008297:Male; D009154:Mutation / Q000235:genetics; D009355:Neomycin; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics; D011995:Recombination, Genetic / Q000187:drug effects; D014162:Transfection / Q000235:genetics / Q000379:methods; D014407:Tumor Cells, Cultured
D011994:Recombinant Proteins; D007041:Hypoxanthine Phosphoribosyltransferase; D017853:Phosphotransferases (Alcohol Group Acceptor); D009355:Neomycin