lorena postiglione, m. biomedical eng. tutor: dr. diego di bernardo xxix cycle – 1 st year...
TRANSCRIPT
Lorena Postiglione, M. Biomedical Eng.Tutor: Dr. Diego di Bernardo
XXIX Cycle – 1st year presentation
Towards Microfluidics-based Automatic Control of gene expression in mammalian cells
Lorena Postiglione 2
Control Engineering in Biology
Aim: To force a population of living cells to produce a desired (fixed or time Aim: To force a population of living cells to produce a desired (fixed or time varying) amount of a proteinvarying) amount of a protein
DESIRED fluorescence
EXPERIMENTAL fluorescence
tetracycline
Microfluidic device and input administration
Martin Kolnik et al., Lab on a Chip - 2012Martin Kolnik et al., Lab on a Chip - 2012
Inducer Input
Untreated Input
SyringeTreated medium
SyringeUntreated medium
Lorena Postiglione 3
Lorena Postiglione 4
Experimental Platform
tetracycline
treated or untreated media
Lorena Postiglione 5
Target reference: 50% of initial steady state fluorescenceCalibration time: 3 hrsControl duration: 3500 minTetracycline @100ng/ml
Set point control: In vitro control
Relay controller PI-PWM controller
Lorena Postiglione 6
Conclusions & Future Plans
• Develop model-based controllers
• Perform tracking control experiment
• Develop a single cell control strategy
Didactics Seminars Research activity
1st year CFU 26 5.4 30
A feedback control strategy can successfully regulate the gene expression in mammalian cells.
Relay and PI controllers don’t achieve optimal performances
Automatic control of gene expression in a mammalian cell lineChiara Fracassi, Lorena Postiglione, Gianfranco Fiore, and Diego di Bernardo, ACS Synthetic Biology (in preparation)
The Second International Mammalian Synthetic Biology Workshop, April 2015,Massachusetts Institute of Technology Cambridge, MA, USA
Lorena Postiglione 7
Acknowledgements
Systems and Synthetic Biology Lab
Lorena Postiglione 8
Lorena Postiglione 9
An algorithm computes a mask on the phase contrast image
Ts =15 min
Image acquisition and real-time processing
Controller design: Relay controller with hysteresis
uer + systemyp
-
In silico relay control
Lorena Postiglione 10
Controller design:Proportional-Integral (PI) controller with Pulse Width Modulator (PWM)
inputuerPI
+ PWM systemyp
-
Lorena Postiglione 11
Controller design:Proportional-Integral (PI) controller with Pulse Width Modulator (PWM)
inputuerPI+ system
yp
-PWM
In silico PI-PWM control
Lorena Postiglione 12
13