SPARKinfect · SPARKinfect: Cracking the Electrical Code of Staphylococcus aureus to Understand How Superbugs Survive in Different Environments

The emergence of multidrug-resistant pathogens such as Staphylococcus aureus (MRSA) poses a global health threat. A central yet underexplored driver of bacterial survival is the membrane potential (Δψ), a fundamental electrical property that regulates energy transduction, cell division, and antibiotic susceptibility. Evidence indicates that Δψ is not a passive backdrop but an active regulator of physiology, enabling adaptation to stress. Yet no systematic framework quantifies or explains Δψ dynamics of S. aureus during infection.SPARKinfect will focus on S. aureus bioelectric physiology during intracellular infection, where bacteria are shielded from antibiotics and immunity. Using Nernstian dyes and confocal–FLIM microscopy, I will build quantitative single-cell maps of Δψ in host cells with high temporal and spatial resolution. Comparative measurements in planktonic cultures and biofilms will serve as reference states to identify infection-specific electrical signatures. I will combine these data with host–pathogen co-culture, and antibiotic challenges to test how electrical states govern persistence and treatment outcomes.The project will first establish how intracellular Δψ dynamics differ from those observed in extracellular lifestyles, then uncover the genetic determinants of electrophysiological adaptation using imaging of mutant libraries, and finally demonstrate how specific electrical states causally shape antibiotic uptake, activity, and survival outcomes. By linking single-cell physiology to infection and treatment, SPARKinfect will provide the first mechanistic framework to explain how Δψ underpins the persistence of S. aureus inside host cells.The results will pioneer a bioelectric perspective on pathogenesis. Beyond basic science, SPARKinfect will yield insights into intracellular antibiotic tolerance, guide novel therapeutic strategies, and establish a versatile pipeline for microbial electrophysiology applicable to other priority pathogens.