Real-time RNA profiling within a single bacterium Journal Article

Author(s): Le, Thuc T.; Harlepp, Sébastien; Guet, Cǎlin C; Dittmar, Kimberly; Emonet, Thierry; Pan, Tao; Cluzel, Philippe
Article Title: Real-time RNA profiling within a single bacterium
Abstract: Characterizing the dynamics of specific RNA levels requires real-time RNA profiling in a single cell. We show that the combination of a synthetic modular genetic system with fluorescence correlation spectroscopy allows us to directly measure in real time the activity of any specific promoter in prokaryotes. Using a simple inducible gene expression system, we found that induced RNA levels within a single bacterium of Escherichia coli exhibited a pulsating profile in response to a steady input of inducer. The genetic deletion of an efflux pump system, a key determinant of antibiotic resistance, altered the pulsating transcriptional dynamics and caused overexpression of induced RNA. In contrast with population measurements, real-time RNA profiling permits identifying relationships between genotypes and transcriptional dynamics that are accessible only at the level of the single cell.
Journal Title: PNAS
Volume: 102
Issue 26
ISSN: 1091-6490
Publisher: National Academy of Sciences  
Date Published: 2005-01-01
Start Page: 9160
End Page: 9164
DOI: 10.1073/pnas.0503311102
Notes: 4237
Open access: no