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Prototyping Diverse Synthetic Biological Circuits in a Cell-Free Transcription-Translation System


Guo, Shaobin (2017) Prototyping Diverse Synthetic Biological Circuits in a Cell-Free Transcription-Translation System. Dissertation (Ph.D.), California Institute of Technology. doi:10.7907/Z9CR5RDK.


Synthetic biological circuits are the foundation for the ultimate goals of controlling cells and building artificial cells from the ground up. To get closer to these goals in a more efficient way, we utilize a cell-free transcription-translation system to help perfect biological circuits for the simplicity, freedom, and convenience that the system offers. In this thesis, we demonstrate three distinct aspects of biological circuits in a cell-free transcription-translation system: circuit dynamics, phosphorylation, and membrane proteins. We start with a simple feedforward circuit, which shows dynamic responses to the input. We first prototype the feedforward circuit in the cell-free system with the aid of mathematical modeling. Then, based on the knowledge learned from prototyping, we successfully implement the circuit in cells. Not only do we show that a circuit with dynamics can be prototyped in the cell- free system, but we also test a more complicated circuit involving a phosphorylation cycle. The phosphorylation-based insulator circuit is prototyped and then a model created for the circuit is shown to be identifiable in the cell-free system. To further expand the capability of the cell-free system, we demonstrate that biologically active membrane proteins can be generated in the cell-free system with engineering, suggesting that even biological circuits requiring membrane proteins can be prototyped in the system. These results help advance our knowledge of both biological circuits and the cell-free transcription-translation system, and bring us one step closer to our ultimate goals of implementing control theory in synthetic biology.

Item Type:Thesis (Dissertation (Ph.D.))
Subject Keywords:TX-TL; cell-free; biocircuits; synthetic biology
Degree Grantor:California Institute of Technology
Division:Chemistry and Chemical Engineering
Major Option:Biochemistry and Molecular Biophysics
Thesis Availability:Public (worldwide access)
Research Advisor(s):
  • Murray, Richard M.
Thesis Committee:
  • Elowitz, Michael B. (chair)
  • Murray, Richard M.
  • Goentoro, Lea A.
  • Sternberg, Paul W.
Defense Date:10 May 2017
Funding AgencyGrant Number
Office of Naval ResearchN00014-13-1-0074
Record Number:CaltechTHESIS:03082017-163613964
Persistent URL:
Related URLs:
URLURL TypeDescription adapted for Ch. 1 adapted for Ch. 2 adapted for Ch. 3
Guo, Shaobin0000-0001-9736-4078
Default Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:10091
Deposited By: Shaobin Guo
Deposited On:05 Jun 2017 23:33
Last Modified:04 Oct 2019 00:15

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