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High-throughput DNA sequencing combined with tridimensional eukaryotic cell culture applied to the study of bacterial pathogenesis: the case of Methicillin-Resistant Staphylococcus aureus


In recent years, the importance of food-producing animals as an expanding reservoir of methicillin-resistant strains of Staphylococcus aureus (MRSA) has attracted worldwide attention. There are indications that these livestock-associated MRSA strains have originated from methicillin-sensitive strains from humans that, during the jump, have lost some well-known phage-carried human virulence genes and acquired antimicrobial resistance genes. Studies that may improve our understanding of the functions and mechanisms of action of the virulence determinants involved in the pathogenesis of MRSA are important for identifying potential intervention targets to prevent or treat the clinical onset of staphylococcal diseases. In this project, high-throughput transposon sequencing will be initiated to study putative virulence genes of the methicillin-resistant Staphylococcus aureus SABRC13, isolated from a Brazilian dairy. Mutants will be screened for virulence changes in a tridimensional model of eukaryotic cell culture. The Swedish team is highly skilled in molecular techniques and large dataset analysis, which will be complemented by the ability of the Brazilian team to obtain 3D eukaryotic cell culture for studies of bacterial pathogens. Together, this constitutes a robust model for large-scale genetic studies of virulence mechanisms in selected pathogens, which will contribute to detecting new targets to interfere with bacterial infection. (AU)

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