In-Vivo Real-Time Control of Protein Expression from Endogenous and Synthetic Gene Networks
Name
Menolascina-2014-In-Vivo Real-Time Co.pdf
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2.32 MB
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Adobe PDF
Checksum (MD5)
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Author(s)
Menolascina, Filippo
Fiore, Gianfranco
Orabona, Emanuele
De Stefano, Luca
Ferry, Mike
Hasty, Jeff
di Bernardo, Mario
di Bernardo, Diego
Date Issued
May 2014
Journal
PLoS Computational Biology
Publisher
Public Library of Science
Citation
Menolascina, Filippo, Gianfranco Fiore, Emanuele Orabona, Luca De Stefano, Mike Ferry, Jeff Hasty, Mario di Bernardo, and Diego di Bernardo. “In-Vivo Real-Time Control of Protein Expression from Endogenous and Synthetic Gene Networks.” Edited by Feilim Mac Gabhann. PLoS Comput Biol 10, no. 5 (May 15, 2014): e1003625.
Version
Final published version
Abstract
We describe an innovative experimental and computational approach to control the expression of a protein in a population of yeast cells. We designed a simple control algorithm to automatically regulate the administration of inducer molecules to the cells by comparing the actual protein expression level in the cell population with the desired expression level. We then built an automated platform based on a microfluidic device, a time-lapse microscopy apparatus, and a set of motorized syringes, all controlled by a computer. We tested the platform to force yeast cells to express a desired fixed, or time-varying, amount of a reporter protein over thousands of minutes. The computer automatically switched the type of sugar administered to the cells, its concentration and its duration, according to the control algorithm. Our approach can be used to control expression of any protein, fused to a fluorescent reporter, provided that an external molecule known to (indirectly) affect its promoter activity is available.
MIT Department
Massachusetts Institute of Technology. Department of Civil and Environmental Engineering
Terms of Use
Creative Commons Attribution
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DOI of Published Version
https://doi.org/10.1371/journal.pcbi.1003625
