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

TitleDevelopment of Persister-FACSeq: a method to massively parallelize quantification of persister physiology and its heterogeneity.
Publication TypeJournal Article
Year of Publication2016
AuthorsHenry, TC, Brynildsen, MP
JournalSci Rep
Volume6
Pagination25100
Date Published2016 05 04
ISSN2045-2322
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.

DOI10.1038/srep25100
Alternate JournalSci Rep
PubMed ID27142337
PubMed Central IDPMC4855238
Grant ListF30 AI114163 / AI / NIAID NIH HHS / United States
R21 AI117009 / AI / NIAID NIH HHS / United States