title: Development of Persister-FACSeq: a method to massively parallelize quantification of persister physiology and its heterogeneity.
abstract: Bacterial persisters are thought to underlie the relapse of chronic infections. Knowledge of persister physiology would illuminate avenues for therapeutic intervention; however, such knowledge has remained elusive because persisters have yet to be segregated from other cell types to sufficient purity. This technical hurdle has stymied progress toward understanding persistence. Here we developed Persister-FACSeq, which is a method that uses fluorescence-activated cell sorting, antibiotic tolerance assays, and next generation sequencing to interrogate persister physiology and its heterogeneity. As a proof-of-concept, we used Persister-FACSeq on a library of reporters to study gene expression distributions in non-growing Escherichia coli, and found that persistence to ofloxacin is inversely correlated with the capacity of non-growing cells to synthesize protein. Since Persister-FACSeq can be applied to study persistence to any antibiotic in any environment for any bacteria that can harbor a fluorescent reporter, we anticipate that it will yield unprecedented knowledge of this detrimental phenotype.
authors.affiliation: Department of Molecular Biology, Princeton University, Princeton, NJ, 08544, USA.|Rutgers Robert Wood Johnson Medical School, Piscataway, NJ, 08854, USA.
authors.forename: Theresa C
authors.initials: TC
authors.lastname: Henry
authors.affiliation: Department of Molecular Biology, Princeton University, Princeton, NJ, 08544, USA.|Department of Chemical and Biological Engineering, Princeton University, Princeton, NJ, 08544, USA.
authors.forename: Mark P
authors.initials: MP
authors.lastname: Brynildsen
mesh_terms: D001424:Bacterial Infections / Q000382:microbiology*; D001431:Bacteriological Techniques / Q000379:methods*; D004361:Drug Tolerance; D004926:Escherichia coli / Q000302:isolation & purification / Q000502:physiology; D005434:Flow Cytometry / Q000379:methods*; D020869:Gene Expression Profiling; D059014:High-Throughput Nucleotide Sequencing