Amphotropic retroviruses with a hybrid long terminal repeat as a tool for gene therapy of cystic fibrosis

Abstract

We have made two retroviral vectors encoding the bacterial beta-galactosidase (lacZ) as a marker gene and a long terminal repeat (LTR) containing an enhancer of the polyoma F101 virus [symbol: see text]. One vector, [symbol: see text], can be used as a test vector in grafting, lineage analysis and gene therapy studies. The other, [symbol: see text] carries an additional unique cloning site in which a gene of interest can be cloned. Titration experiments showed that in human epithelial cell lines, [symbol: see text] produces a transcriptionally active integration more often than the commonly used BAG vector with the wild type LTR. Human epithelial cells in primary culture could be successfully infected. Our data suggest that gene therapy protocols requiring infection in situ, such as in the case of cystic fibrosis, will be hampered by the relatively low local titres that can be achieved at present.