Increased titer of recombinant AAV vectors by gene transfer with adenovirus coupled to DNA-polylysine complexes.
One of the principal problems with using recombinant adeno-associated virus vectors (rAAV) as vehicles for gene delivery has been the difficulty in obtaining high-titer virus stocks after the initial transfection into producer cells. In this report we describe a method for transfecting cells at extremely high efficiency with rAAV vector DNA and complementation plasmid while simultaneously infecting those cells with replication competent adenovirus using adenovirus-polylysine-DNA complexes. We further show that this technique results in an increase in rAAV transducing titer by two orders of magnitude over what is typically achieved by standard calcium phosphate transfection.
Mamounas
M
M
Department of Medicine, University of California, San Diego, La Jolla 92093-0665, USA.
Leavitt
M
M
Yu
M
M
Wong-Staal
F
F
D000256:Adenoviridae / Q000235:genetics* / Q000502:physiology; D004279:DNA, Viral / Q000008:administration & dosage / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics*; D004337:Drug Carriers; D005816:Genetic Complementation Test; D005822:Genetic Vectors; D015497:HIV-1 / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D010957:Plasmids; D011107:Polylysine / Q000008:administration & dosage; D016337:RNA, Catalytic / Q000096:biosynthesis* / Q000235:genetics; D011995:Recombination, Genetic; D014162:Transfection / Q000379:methods*; D014779:Virus Replication
D004279:DNA, Viral; D004337:Drug Carriers; D016337:RNA, Catalytic; D011107:Polylysine