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- W5003167 abstract "Recombinant human adeno-associated virus 2 (AAV) virions were constructed containing a gene for resistance to neomycin (neoR), under the control of either the herpesvirus thymidine kinase (TK) promoter (vTK-Neo), the murine colony-stimulating factor-1 (CSF-1) promoter (vCSF1-Neo) or the CSF-1 promoter plus an upstream human erythroid cell-specific enhancer, HS-2 (vHS2-CSF1-Neo). Recombinant virions were used to infect low-density murine primary bone marrow cells. In hematopoietic progenitor cell assays initiated with cells infected with these recombinant virions, myeloid as well as erythroid cell colonies resistant to the drug G418, a neomycin analogue, were readily obtained, indicating that the murine hematopoietic progenitor cells were susceptible to infection by the recombinant AAV virions and that the transduced neo gene was functionally active in these cells. Whereas only approximately 10% of the colony-forming unit-granulocyte/macrophage (CFU-GM) colonies cloned from mock-infected cells survived the G418-selection at a final active concentration of 250 micrograms/mL of the drug, the extent of the CFU-GM colony formation initiated with the recombinant AAV-Neo virions was as follows: 15% with vTK-Neo, 22% with vCSF1-Neo and 49% with vHS2-CSF1-Neo. In addition, only 14% of the burst-forming unit-erythroid (BFU-E) colonies from mock-infected cells were resistant to G418, whereas 82% of the BFU-E colonies initiated with cells infected with vHS2-CSF1-Neo virions survived the drug selection, suggesting that a human erythroid cell-specific enhancer was able to potentiate expression of the transduced neoR gene from a murine promoter. Individual CFU-GM and BFU-E colonies from mock-infected or recombinant AAV-Neo virus-infected cultures were subjected to polymerase chain reaction (PCR) analysis using a neo-specific synthetic oligonucleotide primer-pair. A 276 bp DNA fragment that hybridized with a neo-specific DNA probe on Southern blots was detected only in colonies cloned from the recombinant virus-infected cells, indicating stable integration of the transduced neo gene. These studies suggest the feasibility of using the AAV-based vector system in an animal model as a prelude to evaluating its safety and efficacy in human gene therapy." @default.
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- W5003167 date "1993-07-01" @default.
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- W5003167 title "Adeno-associated virus 2-mediated gene transfer in murine hematopoietic progenitor cells." @default.
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