TY - JOUR
T1 - In-Vivo Real-Time Control of Protein Expression from Endogenous and Synthetic Gene Networks
AU - Menolascina, Filippo
AU - Fiore, Gianfranco
AU - Orabona, Emanuele
AU - De Stefano, Luca
AU - Ferry, Mike
AU - Hasty, Jeff
AU - di Bernardo, Mario
AU - di Bernardo, Diego
PY - 2014/5/15
Y1 - 2014/5/15
N2 - 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.
AB - 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.
KW - synthetic biology
UR - http://www.scopus.com/inward/record.url?scp=84901649750&partnerID=8YFLogxK
U2 - 10.1371/journal.pcbi.1003625
DO - 10.1371/journal.pcbi.1003625
M3 - Article (Academic Journal)
C2 - 24831205
AN - SCOPUS:84901649750
SN - 1553-734X
VL - 10
JO - PLoS Computational Biology
JF - PLoS Computational Biology
IS - 5
M1 - e1003625
ER -