Development of Persister-FACSeq: a method to massively parallelize quantification of persister physiology and its heterogeneity
| Autor: | Mark P. Brynildsen, Theresa C. Henry |
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| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Genetics Bacteriological Techniques Multidisciplinary Fluorescent reporter Multidrug tolerance Extramural Gene Expression Profiling 030106 microbiology Physiology High-Throughput Nucleotide Sequencing Bacterial Infections Drug Tolerance Cell sorting Biology Flow Cytometry Phenotype Article Gene expression profiling 03 medical and health sciences 030104 developmental biology Escherichia coli |
| Zdroj: | Scientific Reports |
| ISSN: | 2045-2322 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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