Abstract / Description of output
We describe an innovative experimental approach, and a proof of principle investigation, for the application of System Identification techniques to derive quantitative dynamical models of transcriptional regulation in living cells. Specifically, we constructed an experimental platform for System Identification based on a microfluidic device, a time-lapse microscope, and a set of automated syringes all controlled by a computer. The platform allows delivering a time-varying concentration of any molecule of interest to the cells trapped in the microfluidics device (input) and real-time monitoring of a fluorescent reporter protein (output) at a high sampling rate. We tested this platform on the GAL1 promoter in the yeast Saccharomyces cerevisiae driving expression of a green fluorescent protein (Gfp) fused to the GAL1 gene. We demonstrated that the System Identification platform enables accurate measurements of the input (sugars concentrations in the medium) and output (Gfp fluorescence intensity) signals, thus making it possible to apply System Identification techniques to obtain a quantitative dynamical model of the promoter. We explored and compared linear and nonlinear model structures in order to select the most appropriate to derive a quantitative model of the promoter dynamics. Our platform can be used to quickly obtain quantitative models of eukaryotic promoters, currently a complex and time-consuming process.
Original language | English |
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Pages (from-to) | 025106 |
Journal | Chaos: An Interdisciplinary Journal of Nonlinear Science |
Volume | 23 |
Issue number | 2 |
DOIs | |
Publication status | Published - Jun 2013 |
Keywords / Materials (for Non-textual outputs)
- Fluorescence
- Gene Expression Regulation, Fungal
- Models, Genetic
- Promoter Regions, Genetic
- Reproducibility of Results
- Saccharomyces cerevisiae
- Transcription, Genetic