Stochastic protein expression in individual cells at the single molecule level Export

@article{citeulike:553494, abstract = {In a living cell, gene expression—the transcription of DNA to messenger RNA followed by translation to protein—occurs stochastically, as a consequence of the low copy number of DNA and mRNA molecules involved1, 2, 3, 4, 5, 6. These stochastic events of protein production are difficult to observe directly with measurements on large ensembles of cells owing to lack of synchronization among cells. Measurements so far on single cells lack the sensitivity to resolve individual events of protein production. Here we demonstrate a microfluidic-based assay that allows real-time observation of the expression of -galactosidase in living Escherichia coli cells with single molecule sensitivity. We observe that protein production occurs in bursts, with the number of molecules per burst following an exponential distribution. We show that the two key parameters of protein expression—the burst size and frequency—can be either determined directly from real-time monitoring of protein production or extracted from a measurement of the steady-state copy number distribution in a population of cells. Application of this assay to probe gene expression in individual budding yeast and mouse embryonic stem cells demonstrates its generality. Many important proteins are expressed at low levels7, 8, and are thus inaccessible by current genomic and proteomic techniques. This microfluidic single cell assay opens up possibilities for system-wide characterization of the expression of these low copy number proteins.}, author = {Cai, Long and Friedman, Nir and Xie, X. Sunney}, citeulike-article-id = {553494}, citeulike-linkout-0 = {http://dx.doi.org/10.1038/nature04599}, citeulike-linkout-1 = {http://dx.doi.org/10.1038/nature04599}, citeulike-linkout-2 = {http://view.ncbi.nlm.nih.gov/pubmed/16541077}, citeulike-linkout-3 = {http://www.hubmed.org/display.cgi?uids=16541077}, day = {16}, doi = {10.1038/nature04599}, issn = {0028-0836}, journal = {Nature}, keywords = {gene\_expression, noise}, month = {March}, number = {7082}, pages = {358--362}, posted-at = {2006-05-30 00:46:19}, priority = {2}, publisher = {Nature Publishing Group}, title = {Stochastic protein expression in individual cells at the single molecule level}, url = {http://dx.doi.org/10.1038/nature04599}, volume = {440}, year = {2006} }

TY - JOUR ID - citeulike:553494 L3 - citeulike-article-id:553494 N2 - In a living cell, gene expression—the transcription of DNA to messenger RNA followed by translation to protein—occurs stochastically, as a consequence of the low copy number of DNA and mRNA molecules involved1, 2, 3, 4, 5, 6. These stochastic events of protein production are difficult to observe directly with measurements on large ensembles of cells owing to lack of synchronization among cells. Measurements so far on single cells lack the sensitivity to resolve individual events of protein production. Here we demonstrate a microfluidic-based assay that allows real-time observation of the expression of -galactosidase in living Escherichia coli cells with single molecule sensitivity. We observe that protein production occurs in bursts, with the number of molecules per burst following an exponential distribution. We show that the two key parameters of protein expression—the burst size and frequency—can be either determined directly from real-time monitoring of protein production or extracted from a measurement of the steady-state copy number distribution in a population of cells. Application of this assay to probe gene expression in individual budding yeast and mouse embryonic stem cells demonstrates its generality. Many important proteins are expressed at low levels7, 8, and are thus inaccessible by current genomic and proteomic techniques. This microfluidic single cell assay opens up possibilities for system-wide characterization of the expression of these low copy number proteins. IS - 7082 JF - Nature SN - 0028-0836 EP - 362 TI - Stochastic protein expression in individual cells at the single molecule level PB - Nature Publishing Group VL - 440 SP - 358 KW - gene_expression KW - noise AU - Cai, Long AU - Friedman, Nir AU - Xie, Sunney PY - 2006/03/16/ UR - http://dx.doi.org/10.1038/nature04599 ER -