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In vivo gene regulation in Salmonella spp. by a salicylate-dependent control circuit

AuthorsRoyo, José Luis ; Becker, Pablo Daniel; Camacho, Eva María ; Cebolla, Ángel; Link, Claudia; Santero, Eduardo ; Guzmán, Carlos Alberto
KeywordsSalmonella infections
Bacterial proteins
Genetic engineering
Hela cells
Sodium salicylate
3-Phosphoshikimate 1-Carboxyvinyltransferase
Issue Date7-Oct-2007
PublisherNature Publishing Group
CitationNature Methods 4(11): 937-942 (2007)
AbstractSystems allowing tightly regulated expression of prokaryotic genes in vivo are important for performing functional studies of bacterial genes in host-pathogen interactions and establishing bacteria-based therapies. We integrated a regulatory control circuit activated by acetyl salicylic acid (ASA) in attenuated Salmonella enterica that carries an expression module with a gene of interest under control of the XylS2-dependent Pm promoter. This resulted in 20-150-fold induction ex vivo. The regulatory circuit was also efficiently induced by ASA when the bacteria resided in eukaryotic cells, both in vitro and in vivo. To validate the circuit, we administered Salmonella spp., carrying an expression module encoding the 5-fluorocytosine-converting enzyme cytosine deaminase in the bacterial chromosome or in a plasmid, to mice with tumors. Induction with ASA before 5-fluorocytosine administration resulted in a significant reduction of tumor growth. These results demonstrate the usefulness of the regulatory control circuit to selectively switch on gene expression during bacterial infection.
Description6 páginas, 4 figuras. Supplementary information is available on the Nature Methods website.
Publisher version (URL)http://dx.doi.org/10.1038/nmeth1107
Appears in Collections:(CABD) Artículos
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