SV40 As an Effective Gene Transfer Vector in Vivo

SV40 was used to transduce gene expression in vitro and in vivo. Using cloned SV40 genome, we replaced large T antigen gene (Tag) with a polylinker, and inserted firefly luciferase, controlled by SV40 early promoter. Transfection into Tag-expressing cells yielded Tag-deficient virus, SVluc. SVluc was Tag-deficient and therefore replication-deficient in cells that did not supply Tag. SVluc transduced functional luciferase expression in vitro. BALB/c mice were inoculated with SVluc, and their tissues were assayed 3-21 days post-inoculation (dpi) for luciferase protein production and enzyme activity. Luciferase protein was detected by immunohistochemistry throughout the experiment, from 3 to 21 dpi. There was no inflammatory reaction against SVluc-infected cells at any time, in any tissue studied. Luciferase activity was first detected by luminometry 14 dpi, and remained level through day 21. Thus, replication-deficient recombinant SV40 can mediate gene transfer in vitro and in vivo. SV40 was used to transduce gene expression in vitro and in vivo. Using cloned SV40 genome, we replaced large T antigen gene (Tag) with a polylinker, and inserted firefly luciferase, controlled by SV40 early promoter. Transfection into Tag-expressing cells yielded Tag-deficient virus, SVluc. SVluc was Tag-deficient and therefore replication-deficient in cells that did not supply Tag. SVluc transduced functional luciferase expression in vitro. BALB/c mice were inoculated with SVluc, and their tissues were assayed 3-21 days post-inoculation (dpi) for luciferase protein production and enzyme activity. Luciferase protein was detected by immunohistochemistry throughout the experiment, from 3 to 21 dpi. There was no inflammatory reaction against SVluc-infected cells at any time, in any tissue studied. Luciferase activity was first detected by luminometry 14 dpi, and remained level through day 21. Thus, replication-deficient recombinant SV40 can mediate gene transfer in vitro and in vivo.