The invention provides a novel retroviral packaging system, in which
retroviral packaging plasmids and packagable vector transcripts are
produced from high expression plasmids after stable or transient
transfection in mammalian cells. High titers of recombinant retrovirus
are produced in these transfected mammalian cells and can then transduce
a mammalian target cell by cocultivation or supernatant infection. The
methods of the invention include the use of the novel retroviral
packaging plasmids and vectors to transduce primary human cells,
including T cells and human hematopoietic stem cells, with foreign genes
by cocultivation or supernatant infection at high efficiencies. The
invention is useful for the rapid production of high titer viral
supernatants, and to transduce with high efficiency cells that are
refractory to transduction by conventional means.