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dc.contributor.authorRoyo, José Luis
dc.contributor.authorBecker, Pablo Daniel
dc.contributor.authorCamacho, Eva María
dc.contributor.authorCebolla, Angel
dc.contributor.authorLink, Claudia
dc.contributor.authorSantero, Eduardo
dc.contributor.authorGuzmán, Carlos Alberto
dc.date.accessioned2008-04-21T08:57:33Z
dc.date.available2008-04-21T08:57:33Z
dc.date.issued2007-11
dc.identifier.citationIn vivo gene regulation in Salmonella spp. by a salicylate-dependent control circuit. 2007, 4 (11):937-42 Nat. Methodsen
dc.identifier.issn1548-7091
dc.identifier.pmid17922017
dc.identifier.doi10.1038/nmeth1107
dc.identifier.urihttp://hdl.handle.net/10033/23852
dc.description.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.
dc.language.isoenen
dc.subject.mesh3-Phosphoshikimate 1-Carboxyvinyltransferaseen
dc.subject.meshAnimalsen
dc.subject.meshAspirinen
dc.subject.meshBacterial Proteinsen
dc.subject.meshCell Line, Tumoren
dc.subject.meshFlucytosineen
dc.subject.meshFluorouracilen
dc.subject.meshGene Expressionen
dc.subject.meshGene Expression Regulation, Bacterialen
dc.subject.meshGenetic Engineeringen
dc.subject.meshGreen Fluorescent Proteinsen
dc.subject.meshHela Cellsen
dc.subject.meshHumansen
dc.subject.meshLac Operonen
dc.subject.meshMacrophagesen
dc.subject.meshMiceen
dc.subject.meshNeoplasmsen
dc.subject.meshOperonen
dc.subject.meshPromoter Regions (Genetics)en
dc.subject.meshSalmonella Infectionsen
dc.subject.meshSalmonella entericaen
dc.subject.meshSodium Salicylateen
dc.subject.meshSpleenen
dc.subject.meshTranscription Factorsen
dc.subject.meshbeta-Galactosidaseen
dc.titleIn vivo gene regulation in Salmonella spp. by a salicylate-dependent control circuit.en
dc.typeArticleen
dc.contributor.departmentCentro Andaluz de Biología del Desarrollo, Universidad Pablo de Olavide-Consejo Superior de Investigaciones Científicas, Carretera, Utrera, Km 1, E-41013 Sevilla, Spain.en
dc.identifier.journalNature methodsen
refterms.dateFOA2008-06-05T00:00:00Z
html.description.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.


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