Minimally invasive determination of mRNA concentration in single living bacteria. Export

@article{citeulike:2889648, 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 approximately 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.}, address = {The James Franck Institute, Institute for Biophysical Dynamics, Department of Physics, The University of Chicago, 929 E 57th St, Chicago, IL 60637, USA.}, author = {Guet, C\u{a}lin C. and Bruneaux, Luke and Min, Taejin L. and Siegal-Gaskins, Dan and Figueroa, Israel and Emonet, Thierry and Cluzel, Philippe}, citeulike-article-id = {2889648}, citeulike-linkout-0 = {http://dx.doi.org/10.1093/nar/gkn329}, citeulike-linkout-1 = {http://nar.oxfordjournals.org/cgi/content/abstract/36/12/e73}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/18515347}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=18515347}, doi = {10.1093/nar/gkn329}, issn = {1362-4962}, journal = {Nucleic acids research}, keywords = {measurement, mrna}, month = {July}, number = {12}, posted-at = {2008-09-15 09:51:54}, priority = {4}, title = {Minimally invasive determination of mRNA concentration in single living bacteria.}, url = {http://dx.doi.org/10.1093/nar/gkn329}, volume = {36}, year = {2008} }

TY - JOUR ID - citeulike:2889648 L3 - citeulike-article-id:2889648 N2 - 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 approximately 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. IS - 12 JF - Nucleic acids research SN - 1362-4962 AD - The James Franck Institute, Institute for Biophysical Dynamics, Department of Physics, The University of Chicago, 929 E 57th St, Chicago, IL 60637, USA. TI - Minimally invasive determination of mRNA concentration in single living bacteria. VL - 36 KW - measurement KW - mrna AU - Guet, Călin AU - Bruneaux, Luke AU - Min, Taejin AU - Siegal-Gaskins, Dan AU - Figueroa, Israel AU - Emonet, Thierry AU - Cluzel, Philippe PY - 2008/07/30/ UR - http://dx.doi.org/10.1093/nar/gkn329 ER -