Staphylococcus aureus is a gram-positive opportunistic pathogen commonly prevalent in both community and hospital settings. The challenge in treating S. aureus infections arises due to its ability to acquire resistance rapidly against antimicrobials used against it as well as its ability to form biofilms. Reportedly, S. aureus biofilms are under a stringent regulation of two-component systems. To date, 16 Two-component regulators are known in S. aureus however, there’s still a dearth in understanding the precise response regulators that account for biofilm formation in this pathogen. Our study focuses on the transcriptional profile of S. aureus isolates taken from patients undergoing antibiotic therapy in a hospital in New Delhi, India. The gene expression profile in planktonic and biofilm mode of growth of seven clinical isolates indicated that hssRS two component system plays a significant role in biofilm mode of growth. MIC of oxacillin was determined for all the isolates. The expression pattern for all 16 response regulators was checked under antibiotic stressed conditions with MIC50 of oxacillin. The comparative analysis under both conditions showed an interesting differential upregulation of hssRS two component system. A deletion mutant of hssR has been constructed using CRISPR. The resistance pattern and biofilm pattern of the mutant are being analyzed. This two-component system has never been implicated before in biofilm formation and holds promise as a potential target to treat drug resistant S. aureus infections.