Roles of Regulatory Small RNAs in Antibiotic Tolerance in Multidrug Resistant Staphylococcus Aureus

My lab has an ongoing interest in how complex genetic traits such as virulence are regulated and selected in bacterial pathogens. Non-coding RNA (ncRNA) regulation has come to the fore with the advent of RNA sequencing and it has been demonstrated that bacterial pathogens produce hundreds of ncRNAs. However, we have a poor understanding of the function of the majority of these RNA species. The functions of bacterial ncRNAs are likely to be exceptionally diverse, and we are using UV-crosslinking and deep sequencing techniques to study these processes and reveal novel mechanisms of gene regulation.

Staphylococcus aureus is  a major human pathogen that is the leading cause of bacteremia, infective endocarditis, and osteomyelitis. The prevalence of multi-drug resistant S. aureus (MRSA) is rising. MRSA infections are treated with a limited number of last-line antibiotics and many infections are not cleared because of intermediate resistance (antibiotic tolerance). Regulatory small RNAs have been shown to respond to antibiotic treatment and are thought to adapt the cell for antibiotic tolerance.

This project will define the regulatory small RNAs required for vancomycin tolerance in MRSA by constructing a library of sRNA-knockdowns using new CRISPRi technologies. Regulatory small RNAs that are identified in this screen will be further characterised by constructing GFP translational fusions and gene deletions, and using RNA immunoprecipitation techniques to identify new regulatory pathways. 

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