Development of Persister-FACSeq: a method to massively parallelize quantification of persister physiology and its heterogeneity

Theresa C. Henry, Mark P. Brynildsen

Research output: Contribution to journalArticle

15 Scopus citations

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.

Original languageEnglish (US)
Article number25100
JournalScientific reports
Volume6
Issue number1
DOIs
StatePublished - Jul 6 2016

All Science Journal Classification (ASJC) codes

  • General

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