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.
Strayer
D S
DS
Department of Pathology, Anatomy and Cell Biology, Jefferson Medical College, Philadelphia, Pennsylvania 19107, USA.
D000818:Animals; D000952:Antigens, Polyomavirus Transforming / Q000235:genetics; D019556:COS Cells; D002460:Cell Line; D003001:Cloning, Molecular; D003673:Defective Viruses / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D008156:Luciferases / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D013539:Simian virus 40 / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D019065:Virus Assembly
D000952:Antigens, Polyomavirus Transforming; D008156:Luciferases