Adeno-associated virus: a vector system for efficient introduction and integration of DNA into a variety of mammalian cell types.
Adeno-associated virus (AAV) is a single-stranded DNA parvovirus that is dependent on adenovirus or herpesvirus for reproductive functions. We describe the construction of recombinant AAV vectors containing the chloramphenicol acetyltransferase gene or the neomycin phosphotransferase gene. These vectors carried their respective genes into a wide variety of cell types, including primary skin fibroblasts and hematopoietic cells. Infection efficiencies varied with cell type and ranged up to 3.0%. Coinfection of two different recombinant viruses was also used to introduce two different sequences simultaneously into a given cell. Finally, methods for obtaining recombinant AAV vectors with minimal contamination of wild-type virus are described. These various attributes of AAV vectors make them a viable DNA transduction system.
Lebkowski
J S
JS
Applied ImmuneSciences, Inc., Division of Molecular Biology, Menlo Park, California 94025.
McNally
M M
MM
Okarma
T B
TB
Lerch
L B
LB
D000818:Animals; D015139:Blotting, Southern; D002213:Capsid / Q000235:genetics; D002478:Cells, Cultured; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004274:DNA, Recombinant; D000229:Dependovirus / Q000235:genetics*; D005347:Fibroblasts; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells; D019868:Kanamycin Kinase; D010770:Phosphotransferases / Q000235:genetics; D015183:Restriction Mapping
D004274:DNA, Recombinant; D015500:Chloramphenicol O-Acetyltransferase; D010770:Phosphotransferases; D019868:Kanamycin Kinase