Minimally invasive determination of mRNA concentration in single living bacteria

C.C. Guet, L. Bruneaux, T. Min, D. Siegal Gaskins, I. Figueroa, T. Emonet, P. Cluzel, Nucleic Acids Research 36 (2008).

Download
No fulltext has been uploaded. References only!

Journal Article | Published
Author
; ; ; ; ; ;
Abstract
Fluorescence correlation spectroscopy (FCS) has permitted the characterization of high concentrations of noncoding RNAs in a single living bacterium. Here, we extend the use of FCS to low concentrations of coding RNAs in single living cells. We genetically fuse a red fluorescent protein (RFP) gene and two binding sites for an RNA-binding protein, whose translated product is the RFP protein alone. Using this construct, we determine in single cells both the absolute [mRNA] concentration and the associated [RFP] expressed from an inducible plasmid. We find that the FCS method allows us to reliably monitor in real-time [mRNA] down to similar to 40 nM (i.e. approximately two transcripts per volume of detection). To validate these measurements, we show that [mRNA] is proportional to the associated expression of the RFP protein. This FCS-based technique establishes a framework for minimally invasive measurements of mRNA concentration in individual living bacteria.
Publishing Year
Date Published
2008-01-01
Journal Title
Nucleic Acids Research
Acknowledgement
PMCID: PMC2475643
Volume
36
Issue
12
IST-REx-ID

Cite this

Guet CC, Bruneaux L, Min T, et al. Minimally invasive determination of mRNA concentration in single living bacteria. Nucleic Acids Research. 2008;36(12). doi:10.1093/nar/gkn329
Guet, C. C., Bruneaux, L., Min, T., Siegal Gaskins, D., Figueroa, I., Emonet, T., & Cluzel, P. (2008). Minimally invasive determination of mRNA concentration in single living bacteria. Nucleic Acids Research, 36(12). https://doi.org/10.1093/nar/gkn329
Guet, Calin C, Luke Bruneaux, Taejin Min, Dan Siegal Gaskins, Israel Figueroa, Thierry Emonet, and Philippe Cluzel. “Minimally Invasive Determination of MRNA Concentration in Single Living Bacteria.” Nucleic Acids Research 36, no. 12 (2008). https://doi.org/10.1093/nar/gkn329.
C. C. Guet et al., “Minimally invasive determination of mRNA concentration in single living bacteria,” Nucleic Acids Research, vol. 36, no. 12, 2008.
Guet CC, Bruneaux L, Min T, Siegal Gaskins D, Figueroa I, Emonet T, Cluzel P. 2008. Minimally invasive determination of mRNA concentration in single living bacteria. Nucleic Acids Research. 36(12).
Guet, Calin C., et al. “Minimally Invasive Determination of MRNA Concentration in Single Living Bacteria.” Nucleic Acids Research, vol. 36, no. 12, Oxford University Press, 2008, doi:10.1093/nar/gkn329.

Export

Marked Publications

Open Data IST Research Explorer

Search this title in

Google Scholar