Mitigating the effects of ribosome limitations on synthetic circuits via high-gain sRNA-mediated negative feedback
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921845798-MIT.pdf
Description
Full printable version
Size
10.43 MB
Format
Adobe PDF
Checksum (MD5)
3df073c9008edd539f7501e8a3fbdc13
Author(s)
Yazbek, John Elias
Advisor(s)
Domitilla Del Vecchio and Timothy K. Lu.
Date Issued
2015
Publisher
Massachusetts Institute of Technology
Abstract
Resource limitations in bacterial cells can present significant hurdles that preclude correct synthetic circuit behavior. In a simple circuit with one constitutively expressed protein and one protein whose expression is inducible, it has been shown that inducing the expression of the second protein causes a significant decrease in the level of the first. In this thesis, we explore the possibility of reducing the effects of resource limitations by adding a high-gain negative feedback loop to one of the circuits. The loop includes an sRNA construct. We explore different implementations of this circuit and model them mechanistically. Furthermore, we begin physically implementing one of the circuit designs by testing intermediate constructs. Finally, we also explore the hypothesis that exogenous circuits on plasmids compete for a pool of resources that is spatially separated from the resources that the genome utilizes. Through our work, we show results that support the spatial separation hypothesis.
Description
Thesis: S.M., Massachusetts Institute of Technology, Department of Biological Engineering, 2015.
Cataloged from PDF version of thesis.
Includes bibliographical references (pages 85-88).
Subjects
Biological Engineering.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
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