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

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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

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Experimental Platform

tetracycline

treated or untreated media

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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

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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

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Acknowledgements

Systems and Synthetic Biology Lab

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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

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Controller design:Proportional-Integral (PI) controller with Pulse Width Modulator (PWM)

inputuerPI

+ PWM systemyp

-

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Controller design:Proportional-Integral (PI) controller with Pulse Width Modulator (PWM)

inputuerPI+ system

yp

-PWM

In silico PI-PWM control

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