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Visualization of infection and gene delivery by Listeria monocytogenes in live animal
Yong A. Yu*1; Ivaylo Gentschev2; Werner Goebel2; Aladar Szalay3
(1) Dept. of Biochemistry; Dept. of Human Anatomy, Loma Linda University, Loma Linda, CA 92350, USA; (2) Lehrstuhl fur Mikrobiologie, University of Wurzburg, 97074 Wurzburg, Germany; (3) Dept. of Biochemistry, Loma Linda University, Loma Linda, CA 92350 *(email@example.com)
L. monocytogenes has been shown to be able to deliver eukaryotic expression vectors into the cytosol of mammalian cells in tissue cultures. To further investigate the delivery of eukaryotic expression vectors via Listeria in live animals, we constructed a new attenuated suicide Listeria strain transformed with a Renilla luciferase-GFP fusion cassette. In parallel experiments, the same vector is used to generate a prokaryotic expression construct using the SOD promoter to monitor bacterial infection in cells and in animals as control. Both strains are injected into the tail veins of nude mice. Cellular and tissue infections of the whole animal as well as individual organs are visualized under a fluorescence stereo microscope. This Listeria based gene delivery system will serve as a model system for DNA vaccination and gene therapy in live animals. To eliminate the risks associated with the possibility of integration of the plasmids DNA into the host genome, we inserted an Artificial Death Switch gene in the plasmid vectors delivered by Listeria. In this way, host cells carrying the insert can be eliminated by oral or intraperitoneal administration of dimerizing drug to trigger apoptosis of the modified cells.
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