UNDERSTANDING AND CONTROLLING THE COLONIZATION CAPACITIES OF PERSISTER CELLS

Antibiotic tolerance is one of the most critical global public health threats of the 21st century. Bacterial cells that are temporarily resistant to high levels of antibiotics are referred to as persisters. Nearly all bacterial cultures contain a small population of persister cells. Persisters are thought to be responsible for recurring chronic infections and drug-resistant mutants. Persisters are hard to identify and characterize, and the shift between persistence and growth is very difficult to identify. The goal of this project is to develop and validate a new, high-throughput screening approach that could make identification more effective. The project will also engage 4th - 8th grade students in learning opportunities and will provide undergraduates and graduate students opportunities to engage in research as well as the outreach activities. The primary research objective is to investigate persistence colonization phenotypes by (1) utilizing a high throughput fluorescence based screen coupled with transposon mutagenesis and genomic sequencing to detect genes that might be important for persistence, (2) validating potential persistence mediators using promoter libraries and fluorescence microscopy, and (3) performing single-gene and combinatorial knockout studies of candidate persistence-mediating genes that might have therapeutic impact, with a potential focus on known reactive oxygen species regulators. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.