Applying RNA-Seq to investigate the transition into a Viable But Non Culturable State (VBNC) for the intracellular pathogen, Francisella tularensis LVS.

Many species of bacteria, under conditions stress or nutrient limitation, enter a state dormancy referred to as viable but non-culturable (VBNC). VBNC bacteria persist in the environment, are difficult detect and identify by many standard laboratory methods can be altered their susceptibility antibiotics. Entry into is often accompanied morphological changes; however, mechanisms underlying this poorly understood. Francisella transitions rapidly spontaneously state, therefore has potential an excellent model organism for study phenomenon. To investigate transcriptome F. tularensis LVS we extracted RNA from culturable cells carried out RNA-Seq analysis using both Long-read Nanopore Illumina Sequencing. Differentially expressed genes (DEGs) were identified DE-Seq pipeline. Over 300 significantly upregulated ~100 down regulated cells. Amongst some involved transport metals small molecules, have also putative transcriptional regulators that may master controllers process. Our data represents first transcriptomic it state. Identifying transition critical step understanding drive will help us factors allow resuscitation these bacteria. This important implications environmental persistence pathogenicity species. (Supported NIH Grant P20GM103434 West Virginia IDeA Network Biomedical Research Excellence)