Generation of Helper-Free Amphotropic Retroviruses That Transduce a Dominant-Acting, Methotrexate-Resistant Dihydrofolate Reductase Gene

作者：A. Dusty Miller, Ming-Fan Law, Inder M. Verma · 发表于：Molecular and Cellular Biology · 年份：1985 · DOI：10.1128/mcb.5.3.431-437.1985 · 被引用次数：128 · 研究领域：Virus-based gene therapy research、CRISPR and Genetic Engineering、Viral gastroenteritis research and epidemiology

We constructed several retroviruses which transduced a mutant dihydrofolate reductase gene that was resistant to methotrexate inhibition and functioned as a dominant selectable marker. The titer of dihydrofolate reductase-transducing virus produced by virus-producing cells could be increased to very high levels by selection of the cells in increasing concentrations of methotrexate. Helper virus-free dihydrofolate reductase-transducing virus was also generated by using a broad-host-range amphotropic retroviral packaging system. Cell lines producing helper-free dihydrofolate reductase-transducing virus with a titer of 4 X 10(6) per ml were generated. These retroviral vectors should have general utility for high-efficiency transduction of genes in cultured cells and in animals.