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dc.contributor.authorCallura, Jarred Matthewen_US
dc.date.accessioned2018-11-07T15:57:13Z
dc.date.issued2012
dc.date.submitted2012
dc.identifier.otherb39007935
dc.identifier.urihttps://hdl.handle.net/2144/32011
dc.descriptionThesis (Ph.D.)--Boston Universityen_US
dc.descriptionPLEASE NOTE: Boston University Libraries did not receive an Authorization To Manage form for this thesis or dissertation. It is therefore not openly accessible, though it may be available by request. If you are the author or principal advisor of this work and would like to request open access for it, please contact us at open-help@bu.edu. Thank you.en_US
dc.description.abstractFor synthetic biology to make a lasting impact on real-world problems, further increases in the complexity of biomolecular devices are required. Currently, there is a shortage of orthogonal parts that can be assembled to construct highly complex circuits and networks. RNA molecules are a popular source for synthetic biology parts, due to the versatility and predictability of RNA structures. Previously, our lab developed the engineered riboregulator, a RNA-based gene expression system. The advantages of synthetic riboregulation include: physiologically relevant protein production, component modularity, leakage minimization, rapid response time, tunable gene expression, and the ability to independently riboregulate multiple genes simultaneously using orthogonal riboregulator variants. We performed two sets of in vivo experiments that illustrate these unique features and developed two, higher order synthetic devices based on orthogonal riboregulation: the programmable kill switch and the genetic switchboard. The in vivo experiments involved tracking the localization of the TonB protein and manipulating the SOS DNA damage repair network. These studies highlight the ability of our riboregulator to reveal new insights into microbial physiology. Addressing mounting biosecurity concerns, the programmable kill switch employs two riboregulator variants, which regulate two lambda phage proteins, to induce cell lysis rapidly and selectively. Only when we co-expressed the phage proteins did cell suicide occur, and the circuit can link cell death to four different biological signals. To construct a genetic switchboard, we further increased the number of riboregulators in use by designing two new variants. We directly tested our switchboard in a biosensing setup that reports on four environmental signals in single cells using four differentiable reporters. Finally, we utilized the genetic switchboard in a proof-of-concept metabolic engineering application. The metabolism switchboard regulates four metabolic enzymes that control carbon flux through three, E. coli glucose utilization pathways, and we measured its impressive performance across the RNA, protein, and metabolome scales. All together, the applications described here showcase the considerable real-world potential of the engineered riboregulator.en_US
dc.language.isoen_US
dc.publisherBoston Universityen_US
dc.titleSynthetic biology applications of engineered riboregulationen_US
dc.typeThesis/Dissertationen_US
dc.description.embargo2031-01-02
etd.degree.nameDoctor of Philosophyen_US
etd.degree.leveldoctoralen_US
etd.degree.disciplineEngineeringen_US
etd.degree.grantorBoston Universityen_US
dc.identifier.barcode11719032088033
dc.identifier.mmsid99176425590001161


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