Le,Thuc T.; Harlepp, Sébastien; Guet, Calin CISTA ; Dittmar,Kimberly; Emonet,Thierry; Pan,Tao; Cluzel,Philippe
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.
9160 - 9164
Le T, Harlepp S, Guet CC, et al. Real-time RNA profiling within a single bacterium. PNAS. 2005;102(26):9160-9164. doi:10.1073/pnas.0503311102
Le, T., Harlepp, S., Guet, C. C., Dittmar, K., Emonet, T., Pan, T., & Cluzel, P. (2005). Real-time RNA profiling within a single bacterium. PNAS. National Academy of Sciences. https://doi.org/10.1073/pnas.0503311102
Le, Thuc, Sébastien Harlepp, Calin C Guet, Kimberly Dittmar, Thierry Emonet, Tao Pan, and Philippe Cluzel. “Real-Time RNA Profiling within a Single Bacterium.” PNAS. National Academy of Sciences, 2005. https://doi.org/10.1073/pnas.0503311102.
T. Le et al., “Real-time RNA profiling within a single bacterium,” PNAS, vol. 102, no. 26. National Academy of Sciences, pp. 9160–9164, 2005.
Le T, Harlepp S, Guet CC, Dittmar K, Emonet T, Pan T, Cluzel P. 2005. Real-time RNA profiling within a single bacterium. PNAS. 102(26), 9160–9164.
Le, Thuc, et al. “Real-Time RNA Profiling within a Single Bacterium.” PNAS, vol. 102, no. 26, National Academy of Sciences, 2005, pp. 9160–64, doi:10.1073/pnas.0503311102.