ASU Electronic Theses and Dissertations
This collection includes most of the ASU Theses and Dissertations from 2011 to present. ASU Theses and Dissertations are available in downloadable PDF format; however, a small percentage of items are under embargo. Information about the dissertations/theses includes degree information, committee members, an abstract, supporting data or media.
In addition to the electronic theses found in the ASU Digital Repository, ASU Theses and Dissertations can be found in the ASU Library Catalog.
Dissertations and Theses granted by Arizona State University are archived and made available through a joint effort of the ASU Graduate College and the ASU Libraries. For more information or questions about this collection contact or visit the Digital Repository ETD Library Guide or contact the ASU Graduate College at gradformat@asu.edu.
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Description
The portability of genetic tools from one organism to another is a cornerstone of synthetic biology. The shared biological language of DNA-to-RNA-to-protein allows for expression of polypeptide chains in phylogenetically distant organisms with little modification. The tools and contexts are diverse, ranging from catalytic RNAs in cell-free systems to bacterial proteins expressed in human cell lines, yet they exhibit an organizing principle: that genes and proteins may be treated as modular units that can be moved from their native organism to a novel one. However, protein behavior is always unpredictable; drop-in functionality is not guaranteed.
My work characterizes how two different classes of tools behave in new contexts and explores methods to improve their functionality: 1. CRISPR/Cas9 in human cells and 2. quorum sensing networks in Escherichia coli.
1. The genome-editing tool CRISPR/Cas9 has facilitated easily targeted, effective, high throughput genome editing. However, Cas9 is a bacterially derived protein and its behavior in the complex microenvironment of the eukaryotic nucleus is not well understood. Using transgenic human cell lines, I found that gene-silencing heterochromatin impacts Cas9’s ability to bind and cut DNA in a site-specific manner and I investigated ways to improve CRISPR/Cas9 function in heterochromatin.
2. Bacteria use quorum sensing to monitor population density and regulate group behaviors such as virulence, motility, and biofilm formation. Homoserine lactone (HSL) quorum sensing networks are of particular interest to synthetic biologists because they can function as “wires” to connect multiple genetic circuits. However, only four of these networks have been widely implemented in engineered systems. I selected ten quorum sensing networks based on their HSL production profiles and confirmed their functionality in E. coli, significantly expanding the quorum sensing toolset available to synthetic biologists.
My work characterizes how two different classes of tools behave in new contexts and explores methods to improve their functionality: 1. CRISPR/Cas9 in human cells and 2. quorum sensing networks in Escherichia coli.
1. The genome-editing tool CRISPR/Cas9 has facilitated easily targeted, effective, high throughput genome editing. However, Cas9 is a bacterially derived protein and its behavior in the complex microenvironment of the eukaryotic nucleus is not well understood. Using transgenic human cell lines, I found that gene-silencing heterochromatin impacts Cas9’s ability to bind and cut DNA in a site-specific manner and I investigated ways to improve CRISPR/Cas9 function in heterochromatin.
2. Bacteria use quorum sensing to monitor population density and regulate group behaviors such as virulence, motility, and biofilm formation. Homoserine lactone (HSL) quorum sensing networks are of particular interest to synthetic biologists because they can function as “wires” to connect multiple genetic circuits. However, only four of these networks have been widely implemented in engineered systems. I selected ten quorum sensing networks based on their HSL production profiles and confirmed their functionality in E. coli, significantly expanding the quorum sensing toolset available to synthetic biologists.
ContributorsDaer, René (Author) / Haynes, Karmella (Thesis advisor) / Brafman, David (Committee member) / Nielsen, David (Committee member) / Kiani, Samira (Committee member) / Arizona State University (Publisher)
Created2017
Description
Lignocellulosic biomass represents a renewable domestic feedstock that can support large-scale biochemical production processes for fuels and specialty chemicals. However, cost-effective conversion of lignocellulosic sugars into valuable chemicals by microorganisms still remains a challenge. Biomass recalcitrance to saccharification, microbial substrate utilization, bioproduct titer toxicity, and toxic chemicals associated with chemical pretreatments are at the center of the bottlenecks limiting further commercialization of lignocellulose conversion. Genetic and metabolic engineering has allowed researchers to manipulate microorganisms to overcome some of these challenges, but new innovative approaches are needed to make the process more commercially viable. Transport proteins represent an underexplored target in genetic engineering that can potentially help to control the input of lignocellulosic substrate and output of products/toxins in microbial biocatalysts. In this work, I characterize and explore the use of transport systems to increase substrate utilization, conserve energy, increase tolerance, and enhance biocatalyst performance.
ContributorsKurgan, Gavin (Author) / Wang, Xuan (Thesis advisor) / Nielsen, David (Committee member) / Misra, Rajeev (Committee member) / Nannenga, Brent (Committee member) / Arizona State University (Publisher)
Created2018
Description
Synthetic gene networks have evolved from simple proof-of-concept circuits to
complex therapy-oriented networks over the past fifteen years. This advancement has
greatly facilitated expansion of the emerging field of synthetic biology. Multistability is a
mechanism that cells use to achieve a discrete number of mutually exclusive states in
response to environmental inputs. However, complex contextual connections of gene
regulatory networks in natural settings often impede the experimental establishment of
the function and dynamics of each specific gene network.
In this work, diverse synthetic gene networks are rationally designed and
constructed using well-characterized biological components to approach the cell fate
determination and state transition dynamics in multistable systems. Results show that
unimodality and bimodality and trimodality can be achieved through manipulation of the
signal and promoter crosstalk in quorum-sensing systems, which enables bacterial cells to
communicate with each other.
Moreover, a synthetic quadrastable circuit is also built and experimentally
demonstrated to have four stable steady states. Experiments, guided by mathematical
modeling predictions, reveal that sequential inductions generate distinct cell fates by
changing the landscape in sequence and hence navigating cells to different final states.
Circuit function depends on the specific protein expression levels in the circuit.
We then establish a protein expression predictor taking into account adjacent
transcriptional regions’ features through construction of ~120 synthetic gene circuits
(operons) in Escherichia coli. The predictor’s utility is further demonstrated in evaluating genes’ relative expression levels in construction of logic gates and tuning gene expressions and nonlinear dynamics of bistable gene networks.
These combined results illustrate applications of synthetic gene networks to
understand the cell fate determination and state transition dynamics in multistable
systems. A protein-expression predictor is also developed to evaluate and tune circuit
dynamics.
complex therapy-oriented networks over the past fifteen years. This advancement has
greatly facilitated expansion of the emerging field of synthetic biology. Multistability is a
mechanism that cells use to achieve a discrete number of mutually exclusive states in
response to environmental inputs. However, complex contextual connections of gene
regulatory networks in natural settings often impede the experimental establishment of
the function and dynamics of each specific gene network.
In this work, diverse synthetic gene networks are rationally designed and
constructed using well-characterized biological components to approach the cell fate
determination and state transition dynamics in multistable systems. Results show that
unimodality and bimodality and trimodality can be achieved through manipulation of the
signal and promoter crosstalk in quorum-sensing systems, which enables bacterial cells to
communicate with each other.
Moreover, a synthetic quadrastable circuit is also built and experimentally
demonstrated to have four stable steady states. Experiments, guided by mathematical
modeling predictions, reveal that sequential inductions generate distinct cell fates by
changing the landscape in sequence and hence navigating cells to different final states.
Circuit function depends on the specific protein expression levels in the circuit.
We then establish a protein expression predictor taking into account adjacent
transcriptional regions’ features through construction of ~120 synthetic gene circuits
(operons) in Escherichia coli. The predictor’s utility is further demonstrated in evaluating genes’ relative expression levels in construction of logic gates and tuning gene expressions and nonlinear dynamics of bistable gene networks.
These combined results illustrate applications of synthetic gene networks to
understand the cell fate determination and state transition dynamics in multistable
systems. A protein-expression predictor is also developed to evaluate and tune circuit
dynamics.
ContributorsWu, Fuqing (Author) / Wang, Xiao (Thesis advisor) / Haynes, Karmella (Committee member) / Marshall, Pamela (Committee member) / Nielsen, David (Committee member) / Brafman, David (Committee member) / Arizona State University (Publisher)
Created2017
Description
Throughout the Southwest, complex geology and physiography concomitant with climatic variability contribute to diverse stream hydrogeomorphologies. Many riparian plant species store their seeds in soil seed banks, and germinate in response to moisture pulses, but the climatic controls of this response are poorly understood. To better understand the ecological implications of a changing climate on riparian plant communities, I investigated seed bank responses to seasonal temperature patterns and to stream hydrogeomorphic type. I asked the following questions: Are there distinct suites of warm and cool temperature germinating species associated with Southwestern streams; how do they differ between riparian and terrestrial zones, and between ephemeral and perennial streams? How does alpha diversity of the soil seed bank differ between streams with ephemeral, intermittent, and perennial flow, and between montane and basin streams? Do streams with greater elevational change have higher riparian zone seed bank beta-diversity? Does nestedness or turnover contribute more to within stream beta-diversity?
I collected soil samples from the riparian and terrestrial zones of 21 sites, placing them in growth chambers at one of two temperature regimes, and monitoring emergence of seedlings for 12 weeks. Results showed an approximately equal number of warm and cool specialists in both riparian and terrestrials zones; generalists also were abundant, particularly in the riparian zone. The number of temperature specialists and generalists in the riparian zones did not differ significantly between perennial headwater and ephemeral stream types. In montane streams, alpha diversity of the soil seed bank was highest for ephemeral reaches; in basin streams the intermittent and perennial reaches had higher diversity. Spatial turnover was primarily responsible for within stream beta-diversity—reaches had different species assemblages. The large portion of temperature specialists found in riparian seed banks indicates that even with available moisture riparian zone plant community composition will likely be impacted by changing temperatures. However, the presence of so many temperature generalists in the riparian zones suggests that some component of the seed bank is adapted to variable conditions and might offer resilience in a changing climate. Study results confirm the importance of conserving multiple hydrogeomorphic reach types because they support unique species assemblages.
I collected soil samples from the riparian and terrestrial zones of 21 sites, placing them in growth chambers at one of two temperature regimes, and monitoring emergence of seedlings for 12 weeks. Results showed an approximately equal number of warm and cool specialists in both riparian and terrestrials zones; generalists also were abundant, particularly in the riparian zone. The number of temperature specialists and generalists in the riparian zones did not differ significantly between perennial headwater and ephemeral stream types. In montane streams, alpha diversity of the soil seed bank was highest for ephemeral reaches; in basin streams the intermittent and perennial reaches had higher diversity. Spatial turnover was primarily responsible for within stream beta-diversity—reaches had different species assemblages. The large portion of temperature specialists found in riparian seed banks indicates that even with available moisture riparian zone plant community composition will likely be impacted by changing temperatures. However, the presence of so many temperature generalists in the riparian zones suggests that some component of the seed bank is adapted to variable conditions and might offer resilience in a changing climate. Study results confirm the importance of conserving multiple hydrogeomorphic reach types because they support unique species assemblages.
ContributorsSetaro, Danika (Author) / Stromberg, Juliet (Thesis advisor) / Franklin, Janet (Committee member) / Makings, Elizabeth (Committee member) / Arizona State University (Publisher)
Created2016
Description
Bouteloua eriopoda (Torr.) Torr., also known as black grama, is a perennial bunchgrass native to arid and semiarid ecosystems in the southwestern region of North America. As a result of anthropogenic climate change, this region is predicted to increase in aridity and experience more frequent extreme drought and extreme wet years. This change in precipitation will no doubt affect black grama; however, few studies have investigated how the specific structural components of this grass will respond. The purpose of this study was to examine the effects of years since start of treatment and annual precipitation amount on tiller and stolon densities, and to test for interaction between the two predictor variables. Additionally, the effects of annual precipitation on ramets and axillary buds were investigated. By using 36 experimental plots that have been receiving drought, irrigated, or control treatments since 2007, tiller density was the most responsive component to both annual precipitation amount and years since start of treatment. Years since start of treatment and annual precipitation amount also had a statistically significant interaction, meaning the effect of precipitation amount on tiller density differs depending on how many years have passed since treatments began. Stolon density was the second-most responsive component; the predictor variables were found to have no statistically significant interaction, meaning their effects on stolon density are independent of one another. Ramet density, ramets per stolon, and axillary bud metabolic activity and density were found to be independent of annual precipitation amount for 2021. The results indicate that multiple-year extreme wet and multiple-year extreme dry conditions in the Southwest will both likely reduce tiller and stolon densities in black grama patches. Prolonged drought conditions reduced tiller and stolon production in black grama because of negative legacies from previous years. Reduced production during prolonged wet conditions could be due to increased competition between adjacent plants.
ContributorsSutter, Bryce Madison (Author) / Sala, Osvaldo E (Thesis advisor) / Makings, Elizabeth (Committee member) / Wojciechowski, Martin F (Committee member) / Arizona State University (Publisher)
Created2022