Cotransduction of nondividing cells using lentiviral vectors

Diseases such as AIDS and cancers may require the introduction of multiple genes into either stem cells or nondividing cells, among others, for therap eutic purposes. Such genes may act at different points of the disease pathw ay or may constitute a regulatory loop to bypass or rectify the defective g ene or pathway underpinning the disease. ideally, the therapeutic genes mus t be transduced together in diverse combinations, and the introduction shou ld occur without constraints. Since lentiviral vectors can transduce both d ividing and nondividing cells, they are ideal vehicles to investigate combi natorial gene transfer into diverse cells. In this study, we demonstrate th at by using two independent lentiviral vectors, pseudotyped with the protei n g of vesicular stomatitis virus, up to four genes can be introduced simul taneously into single dividing and nondividing cells. Up to 45% and 73% of dividing and nondividing cells, respectively, could be transduced with two lentiviral vectors. The efficiency of cotransducing a single cell was the p roduct of the individual transduction efficiencies and suggested the absenc e of viral interference. Multiple and combinatorial gene transduction using lentiviral vectors may prove useful in gene therapy.