Nanobody-mediated dynamic and spatial control of cellular proteins for mammalian gene circuit engineering

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dc.contributor.advisorSegatori, Lauraen_US
dc.creatorKuypers, Briannaen_US
dc.date.accessioned2019-11-25T13:52:04Zen_US
dc.date.available2020-06-01T05:01:09Zen_US
dc.date.created2019-12en_US
dc.date.issued2019-11-22en_US
dc.date.submittedDecember 2019en_US
dc.date.updated2019-11-25T13:52:04Zen_US
dc.description.abstractCellular information processing in mammalian cells relies on sophisticated and highly dynamic mechanisms. Unravelling these mechanisms remains a challenging endeavor and has limited de novo design of biological functions. Controlling cellular phenotypes requires precise control of protein concentration and localization, which is typically realized through the design of genetic networks orthogonal to the cellular circuitry. This work describes my efforts to develop tools for achieving temporal and spatial control over cellular proteins through post-translational regulation that can be generally used to characterize and manipulate the complex genetic networks that regulate mammalian cells. I first illustrated the use of a nanobody-based platform designed to control degradation of target proteins (the NanoDeg) to improve the dynamic range and temporal resolution of input-dependent reporter systems. I then explored integration of a reporter-specific NanoDeg into a genetic circuit through a feedforward loop to generate an improved heat-shock reporter system. To enhance temporal control of genetic networks, I investigated strategies to use the NanoDeg for controlling the oscillatory behavior of a range of different circuit topologies. Finally, I developed a nanobody-based platform for controlling the subcellular localization of target proteins. Using this strategy, I experimentally demonstrated selective target localization to multiple cellular compartments and investigated different circuit architectures for dynamic control of target localization to different compartments. Combining nanobody-mediated localization and degradation with orthogonal transcriptional regulation I developed a nanobody-based toolkit to achieve spatial and temporal control of target protein levels.en_US
dc.embargo.terms2020-06-01en_US
dc.format.mimetypeapplication/pdfen_US
dc.identifier.citationKuypers, Brianna. "Nanobody-mediated dynamic and spatial control of cellular proteins for mammalian gene circuit engineering." (2019) Diss., Rice University. <a href="https://hdl.handle.net/1911/107736">https://hdl.handle.net/1911/107736</a>.en_US
dc.identifier.urihttps://hdl.handle.net/1911/107736en_US
dc.language.isoengen_US
dc.rightsCopyright is held by the author, unless otherwise indicated. Permission to reuse, publish, or reproduce the work beyond the bounds of fair use or other exemptions to copyright law must be obtained from the copyright holder.en_US
dc.subjectnanobodyen_US
dc.subjectmammalian gene circuiten_US
dc.subjectdegradationen_US
dc.subjectlocalizationen_US
dc.subjectorthogonal regulationen_US
dc.subjectpost-translational protein regulationen_US
dc.titleNanobody-mediated dynamic and spatial control of cellular proteins for mammalian gene circuit engineeringen_US
dc.typeThesisen_US
dc.type.materialTexten_US
thesis.degree.departmentSystems, Synthetic and Physical Biologyen_US
thesis.degree.disciplineNatural Sciencesen_US
thesis.degree.grantorRice Universityen_US
thesis.degree.levelDoctoralen_US
thesis.degree.majorSynthetic Biologyen_US
thesis.degree.nameDoctor of Philosophyen_US
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