{"abstract":[{"text":"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.","lang":"eng"}],"status":"public","date_published":"2008-01-01T00:00:00Z","acknowledgement":"PMCID: PMC2475643 ","publist_id":"2474","license":"https://creativecommons.org/licenses/by-nc/4.0/","tmp":{"image":"/images/cc_by_nc.png","legal_code_url":"https://creativecommons.org/licenses/by-nc/4.0/legalcode","short":"CC BY-NC (4.0)","name":"Creative Commons Attribution-NonCommercial 4.0 International (CC BY-NC 4.0)"},"type":"journal_article","publication_status":"published","extern":1,"volume":36,"publisher":"Oxford University Press","citation":{"ieee":"C. C. Guet <i>et al.</i>, “Minimally invasive determination of mRNA concentration in single living bacteria,” <i>Nucleic Acids Research</i>, vol. 36, no. 12. Oxford University Press, 2008.","short":"C.C. Guet, L. Bruneaux, T. Min, D. Siegal Gaskins, I. Figueroa, T. Emonet, P. Cluzel, Nucleic Acids Research 36 (2008).","ama":"Guet CC, Bruneaux L, Min T, et al. Minimally invasive determination of mRNA concentration in single living bacteria. <i>Nucleic Acids Research</i>. 2008;36(12). doi:<a href=\"https://doi.org/10.1093/nar/gkn329\">10.1093/nar/gkn329</a>","chicago":"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.” <i>Nucleic Acids Research</i>. Oxford University Press, 2008. <a href=\"https://doi.org/10.1093/nar/gkn329\">https://doi.org/10.1093/nar/gkn329</a>.","apa":"Guet, C. C., Bruneaux, L., Min, T., Siegal Gaskins, D., Figueroa, I., Emonet, T., &#38; Cluzel, P. (2008). Minimally invasive determination of mRNA concentration in single living bacteria. <i>Nucleic Acids Research</i>. Oxford University Press. <a href=\"https://doi.org/10.1093/nar/gkn329\">https://doi.org/10.1093/nar/gkn329</a>","mla":"Guet, Calin C., et al. “Minimally Invasive Determination of MRNA Concentration in Single Living Bacteria.” <i>Nucleic Acids Research</i>, vol. 36, no. 12, Oxford University Press, 2008, doi:<a href=\"https://doi.org/10.1093/nar/gkn329\">10.1093/nar/gkn329</a>.","ista":"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)."},"month":"01","publication":"Nucleic Acids Research","issue":"12","_id":"3754","year":"2008","day":"01","doi":"10.1093/nar/gkn329","quality_controlled":0,"intvolume":"        36","author":[{"full_name":"Calin Guet","first_name":"Calin C","last_name":"Guet","orcid":"0000-0001-6220-2052","id":"47F8433E-F248-11E8-B48F-1D18A9856A87"},{"full_name":"Bruneaux,Luke","last_name":"Bruneaux","first_name":"Luke"},{"first_name":"Taejin","last_name":"Min","full_name":"Min,Taejin L"},{"last_name":"Siegal Gaskins","first_name":"Dan","full_name":"Siegal-Gaskins,Dan"},{"first_name":"Israel","last_name":"Figueroa","full_name":"Figueroa,Israel"},{"full_name":"Emonet,Thierry","last_name":"Emonet","first_name":"Thierry"},{"first_name":"Philippe","last_name":"Cluzel","full_name":"Cluzel,Philippe"}],"date_updated":"2021-01-12T07:51:57Z","date_created":"2018-12-11T12:04:59Z","title":"Minimally invasive determination of mRNA concentration in single living bacteria"}