Research: Enterococcus faecalis is a Gram-positive, biofilm-forming pathobiont that is a leading cause of hospital-acquired infections. Protein regulators of E. faecalis biofilm formation are highly studied, but very little is known about the role of noncoding RNAs (ncRNAs) in gene regulation. Transcriptomic studies predict approximately 230 ncRNAs in the vancomycin-resistant E. faecalis strain V583. I used sequence homology with V583 to predict at least 130 ncRNAs in the well-studied plasmid-free strain E. faecalis OG1RF. I then identified transposon insertion mutants for these predicted regulatory RNA elements and created a targeted arrayed library with these mutants. I screened growth and biofilm formation in this mini library across 8 conditions and 2 timepoints to link these regulatory factors to biologically relevant phenotypes. In my thesis work, I aim to decipher the nutrient-responsive functions of sRNA_00048 and characterize the role of ncRNAs in biofilm formation. This will shed light on the understudied ncRNA-based arm of gene regulation in E. faecalis and could potentially inform the development of new treatments against this highly antibiotic-resistant bacterium.
