Isolation and Identification of Outer Membrane Vesicles cargo of Pseudomonas aeruginosa and Its Implications for Biofilm Regulation
Faculty Mentor
Vinaya Sampath
Area of Research
Microbiology, Molecular Biology
Major
Biomedical Sciences
Description
INTRODUCTION: Pseudomonas aeruginosa PAO1 is a Gram-negative opportunistic pathogen known to cause infections, particularly in immunocompromised individuals. The pathogen is known for its robust biofilm-forming capacity and intrinsic resistance to a wide variety of antimicrobial agents. Pseudomonas aeruginosa secretes outer membrane vesicles (OMVs), which are nanoscale phospholipid bilayer structures constitutively released from the outer membrane and have emerged as important mediators of intercellular communication, stress response, and virulence factor delivery. Despite growing interest in OMV biology, not much is known about the nucleic acid cargo encapsulated within or on the surface of PAO1-derived OMVs and their potential role in biofilm regulation.
METHOD: This study aims to identify the nucleic acid content of OMVs isolated from planktonic PAO1 cultures. OMVs were successfully isolated from cell-free supernatants and purified using density gradient centrifugation. The presence of nucleic acid cargo has been confirmed by gel electrophoresis. Subsequent DNA and small RNA extraction and sequencing are underway.
RESULTS: We hypothesize that P. aeruginosa PAO1 selectively encapsulates a subset of DNA and sRNA into the OMVs during its growth, including regulatory sRNA.
DISCUSSION/CONCLUSION: Understanding the DNA and RNA cargo will provide insight into bacterial communication and how these OMVs can be repurposed as targeted delivery vehicles for antimicrobial agents.
Isolation and Identification of Outer Membrane Vesicles cargo of Pseudomonas aeruginosa and Its Implications for Biofilm Regulation
INTRODUCTION: Pseudomonas aeruginosa PAO1 is a Gram-negative opportunistic pathogen known to cause infections, particularly in immunocompromised individuals. The pathogen is known for its robust biofilm-forming capacity and intrinsic resistance to a wide variety of antimicrobial agents. Pseudomonas aeruginosa secretes outer membrane vesicles (OMVs), which are nanoscale phospholipid bilayer structures constitutively released from the outer membrane and have emerged as important mediators of intercellular communication, stress response, and virulence factor delivery. Despite growing interest in OMV biology, not much is known about the nucleic acid cargo encapsulated within or on the surface of PAO1-derived OMVs and their potential role in biofilm regulation.
METHOD: This study aims to identify the nucleic acid content of OMVs isolated from planktonic PAO1 cultures. OMVs were successfully isolated from cell-free supernatants and purified using density gradient centrifugation. The presence of nucleic acid cargo has been confirmed by gel electrophoresis. Subsequent DNA and small RNA extraction and sequencing are underway.
RESULTS: We hypothesize that P. aeruginosa PAO1 selectively encapsulates a subset of DNA and sRNA into the OMVs during its growth, including regulatory sRNA.
DISCUSSION/CONCLUSION: Understanding the DNA and RNA cargo will provide insight into bacterial communication and how these OMVs can be repurposed as targeted delivery vehicles for antimicrobial agents.