@article{3754,
  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.},
  author       = {Guet, Calin C and Bruneaux, Luke and Min, Taejin and Siegal Gaskins, Dan and Figueroa, Israel and Emonet, Thierry and Cluzel, Philippe},
  issn         = {1362-4962},
  journal      = {Nucleic Acids Research},
  number       = {12},
  publisher    = {Oxford University Press},
  title        = {{Minimally invasive determination of mRNA concentration in single living bacteria}},
  doi          = {10.1093/nar/gkn329},
  volume       = {36},
  year         = {2008},
}