Matching Items (5)
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Description
Arsenic contamination in groundwater is a serious problem both in local Arizonan communities and abroad: prolonged exposure to arsenic contamination can cause cancer, vascular damage, and liver failure. This project aims to engineer the microalgae Chlamydomonas reinhardtii to sequester arsenic out of water. Metallothionein, arsenate reductase, and ferritin were integrated

Arsenic contamination in groundwater is a serious problem both in local Arizonan communities and abroad: prolonged exposure to arsenic contamination can cause cancer, vascular damage, and liver failure. This project aims to engineer the microalgae Chlamydomonas reinhardtii to sequester arsenic out of water. Metallothionein, arsenate reductase, and ferritin were integrated into the microalgae via the pASapI plasmid. The plasmid rescues function of the photosystem II gene, leveraging the ability to photosynthesize as a selective trait. Metallothionein and ferritin bind the two most common forms of arsenic: arsenite and arsenate, respectively. Arsenate reductase catalyzes the reduction of arsenate to arsenite, allowing for the ultimate sequestration of the toxic metal to occur in the chloroplast. The algae was transformed using a biolistic device, to create three mutant strains, expressing Metallothionein (MT), Arsenate Reductase (ArsC)-HA, and MT-6xHIS plasmids respectively. When testing the fluorescence output of these three strains, they showed a maximum quantum yield of photosystem II comparable to that of the wildtype algae, indicating that the rescue gene had been incorporated into the chloroplast genome properly. Strains were exposed to arsenic-containing media at 50ppb and 500 ppb for 48 and 72 hours to determine the arsenic sequestration rate. Arsenic concentration in the supernatant was measured using ICP-MS analysis and sequestration rate was calculated in terms of arsenic concentration per fold growth of algae. The normalized arsenic sequestration rates of tagged protein expressing strains at 50 ppb were significantly higher than wildtype.
ContributorsLieberman, Emma (Author) / Bartelle, Benjamin (Thesis director) / Redding, Kevin (Committee member) / Barrett, The Honors College (Contributor) / Harrington Bioengineering Program (Contributor)
Created2022-05
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Description

We describe a secondary analysis of an in vitro experiment that supports the capabilities of a relatively new imaging technique known as functional Magnetic Resonance Electrical Impedance Tomography (fMREIT) to detect conductivity changes in neural tissue caused by activity. Methods: Magnetic Resonance (MR) phase data of active Aplysia ganglia tissue

We describe a secondary analysis of an in vitro experiment that supports the capabilities of a relatively new imaging technique known as functional Magnetic Resonance Electrical Impedance Tomography (fMREIT) to detect conductivity changes in neural tissue caused by activity. Methods: Magnetic Resonance (MR) phase data of active Aplysia ganglia tissue in artificial seawater (ASW) were collected before and after exposure to an excitotoxin using two different imaging current strengths, and these data were then used to reconstruct conductivity changes throughout the tissue. Results: We found that increases in neural activity led to significant increases in imaged conductivity when using high imaging currents, but these differences in conductivity were not seen in regions that did not contain neural tissue nor in data where there were no differences in neural activity. Conclusion: We conclude that the analysis presented here supports fMREIT as a contrast technique capable of imaging neural activity in live tissue more directly than functional imaging methods such as BOLD fMRI.

ContributorsBarnett, Cole (Author) / Sadleir, Rosalind (Thesis director) / Buneo, Christopher (Committee member) / Bartelle, Benjamin (Committee member) / Barrett, The Honors College (Contributor) / Harrington Bioengineering Program (Contributor)
Created2022-05
Description
By 2050, feeding the world will require a 70% increase in food production with fewer water resources due to climate change. New strategies are needed to replace current approaches. C3 photosynthesis is inefficient due to photorespiration, but synthetic biology offers a way to increase photosynthetic efficiency and crop yields, such

By 2050, feeding the world will require a 70% increase in food production with fewer water resources due to climate change. New strategies are needed to replace current approaches. C3 photosynthesis is inefficient due to photorespiration, but synthetic biology offers a way to increase photosynthetic efficiency and crop yields, such as the tartronyl-CoA (TaCo) pathway. This project assesses the TaCo pathway in the chloroplast of Chlamydomonas reinhardtii and represents a pivotal step toward its practical application in higher plants for use in agriculture and biotechnology.
ContributorsCerna, Gabriella (Author) / Sharma, Priyati (Co-author) / Redding, Kevin (Thesis director) / Bartelle, Benjamin (Committee member) / Barrett, The Honors College (Contributor) / School of Life Sciences (Contributor) / School of Molecular Sciences (Contributor)
Created2024-05
Description
By 2050, feeding the world will require a 70% increase in food production with fewer water resources due to climate change. New strategies are needed to replace current approaches. C3 photosynthesis is inefficient due to photorespiration, but synthetic biology offers a way to increase photosynthetic efficiency and crop yields, such

By 2050, feeding the world will require a 70% increase in food production with fewer water resources due to climate change. New strategies are needed to replace current approaches. C3 photosynthesis is inefficient due to photorespiration, but synthetic biology offers a way to increase photosynthetic efficiency and crop yields, such as the tartronyl-CoA (TaCo) pathway. This project assesses the TaCo pathway in the chloroplast of Chlamydomonas reinhardtii and represents a pivotal step toward its practical application in higher plants for use in agriculture and biotechnology.
ContributorsSharma, Priyati (Author) / Cerna, Gabriella (Co-author) / Redding, Kevin (Thesis director) / Bartelle, Benjamin (Committee member) / Erb, Tobias (Committee member) / Barrett, The Honors College (Contributor) / Harrington Bioengineering Program (Contributor)
Created2024-05
Description

Protein and gene circuit level synthetic bioengineering can require years to develop a single target. Phage assisted continuous evolution (PACE) is a powerful new tool for rapidly engineering new genes and proteins, but the method requires an automated cell culture system, making it inaccessible to non industrial research programs. Complex

Protein and gene circuit level synthetic bioengineering can require years to develop a single target. Phage assisted continuous evolution (PACE) is a powerful new tool for rapidly engineering new genes and proteins, but the method requires an automated cell culture system, making it inaccessible to non industrial research programs. Complex protein functions, like specific binding, require similarly dynamic PACE selection that can be alternatively induced or suppressed, with heat labile chemicals like tetracycline. Selection conditions must be controlled continuously over days, with adjustments made every few minutes. To make PACE experiments accessible to the broader community, we designed dedicated cell culture hardware and integrated optogenetically controlled plasmids. The low cost and open source platform allows a user to conduct PACE with continuous monitoring and precise control of evolution using light.

ContributorsTse, Ashley (Author) / Bartelle, Benjamin (Thesis director) / Tian, Xiaojun (Committee member) / Barrett, The Honors College (Contributor) / Materials Science and Engineering Program (Contributor) / School of International Letters and Cultures (Contributor) / Harrington Bioengineering Program (Contributor)
Created2023-05