Dual RNA-seq of intracellular S. aureus pKikume with different metabolic states within macrophages. S. aureus pKikume

The overall aim of this study was to investigate the strategies used by S. aureus to survive and multiply within professional phagocytes (macrophages). For this purpose, an S. aureus strain encoding the reporter mKikumeGR that enables to monitor the metabolic activity and proliferation status of the internalized bacteria was used to simultaneously assess gene expression changes in host cells and pathogens in a dual RNA-seq approach and determine the correlation between these changes and the metabolic state of internalized bacteria. Therefore, macrophages were infected with S. aureus pKikume, sorted via FACS 4h p.I. into one subpopulation of macrophages harboring mainly metabolic active, green bacteria and one population harboring metabolic inactive, red bacteria, and dual RNA-seq was performed on both subpopulations.