Rational design of modular circuits for gene transcription: A test of the bottom-up approach.
BACKGROUND
Most of synthetic circuits developed so far have been designed by an ad hoc approach, using a small number of components (i.e. LacI, TetR) and a trial and error strategy. We are at the point where an increasing number of modular, inter-changeable and well-characterized components is needed to expand the construction of synthetic devices and to allow a rational approach to the design.


RESULTS
We used interchangeable modular biological parts to create a set of novel synthetic devices for controlling gene transcription, and we developed a mathematical model of the modular circuits. Model parameters were identified by experimental measurements from a subset of modular combinations. The model revealed an unexpected feature of the lactose repressor system, i.e. a residual binding affinity for the operator site by induced lactose repressor molecules. Once this residual affinity was taken into account, the model properly reproduced the experimental data from the training set. The parameters identified in the training set allowed the prediction of the behavior of networks not included in the identification procedure.


CONCLUSIONS
This study provides new quantitative evidences that the use of independent and well-characterized biological parts and mathematical modeling, what is called a bottom-up approach to the construction of gene networks, can allow the design of new and different devices re-using the same modular parts.
Laboratory of Cellular and Molecular Engineering, University of Bologna, I-47521 Cesena, Italy.|Department of Electronics, Computer Science and Systems, University of Bologna, I-47521 Cesena, Italy.
Francesca
F
Ceroni
Department of Medical Surgery and Bioengineering, University of Siena, I-53100 Siena, Italy.
Simone
S
Furini
Laboratory of Cellular and Molecular Engineering, University of Bologna, I-47521 Cesena, Italy.|Department of Biochemistry "G. Moruzzi", University of Bologna, I-40126 Bologna, Italy.
Emanuele
E
Giordano
Laboratory of Cellular and Molecular Engineering, University of Bologna, I-47521 Cesena, Italy.|Department of Electronics, Computer Science and Systems, University of Bologna, I-47521 Cesena, Italy.
Silvio
S
Cavalcanti