In-vivo real-time control of protein expression from endogenous and synthetic gene networks.
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.
TeleThon Institute of Genetics and Medicine (TIGEM), Naples, Italy; Department of Electrical Engineering and Information Technology, University of Naples Federico II, Naples, Italy.
Filippo
F
Menolascina
TeleThon Institute of Genetics and Medicine (TIGEM), Naples, Italy; Department of Electrical Engineering and Information Technology, University of Naples Federico II, Naples, Italy.
Gianfranco
G
Fiore
Institute of Microelectronics and Microsystems (IMM), CNR, Naples, Italy.
Emanuele
E
Orabona
Institute of Microelectronics and Microsystems (IMM), CNR, Naples, Italy.
Luca
L
De Stefano
Department of Bioengineering, University of California, San Diego, La Jolla, California, United States of America.
Mike
M
Ferry
Department of Bioengineering, University of California, San Diego, La Jolla, California, United States of America.
Jeff
J
Hasty
Department of Electrical Engineering and Information Technology, University of Naples Federico II, Naples, Italy; Department of Engineering Mathematics, University of Bristol, Bristol, United Kingdom.
Mario
M
di Bernardo
TeleThon Institute of Genetics and Medicine (TIGEM), Naples, Italy; Department of Electrical Engineering and Information Technology, University of Naples Federico II, Naples, Italy.
Diego
D
di Bernardo
D003198:Computer Simulation; D003199:Computer Systems; D005656:Fungal Proteins / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal / Q000235:genetics*; D005813:Genes, Synthetic; D060847:Metabolic Engineering / Q000379:methods*; D008957:Models, Genetic; D015003:Yeasts / Q000502:physiology*
D005656:Fungal Proteins