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            Index:
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          <div class="genTitle">
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            <div>Implementation</div>
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          <div class="sectionTitle" id="Section 1">Testing Efficacy</div>
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                Beyond a proof of concept to determine the success of Manifold, further tests will be required to determine the effectiveness of our design and its scalability. The primary test for efficacy will be an HPLC analysis of the culture broth in order to identify the concentration of resveratrol being produced. To see if the Manifold system adequately increases the flux through the pathway, the final ratios of resveratrol to 4-coumaric acid can be compared. If there is no flux leakage, then this ratio will be equal to 1. In order to see if an increase in overall product is observed, the ratio of resveratrol concentrations between the experimental group and control groups can be used. Finally, to see if the rate of resveratrol is increased by the system, a linear regression can be fit to each concentration vs time plot for resveratrol and the slopes can be compared. If our yields prove that Manifold nearly eliminates flux leakage, we have the data required to move on to testing other enzyme pathways.
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          <div class="sectionTitle" id="Section 2">Further Validating Manifold</div>
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              If our implementation for Manifold proves successful with resveratrol, we can move onto testing other enzyme pathways in similar fashions. Proving Manifold with pathways that are applicable to more important pharmaceutical pathways will be the priority, and we have explored options relating to statins, medications meant to reduce cholesterol levels, in order to extend our system to the production of more diverse compounds. Doing so will demonstrate the scalability of Manifold, and show that our platform technology in the space of biologics has the potential to increase yields and bolster efficiency. 
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          <div class="sectionTitle" id="Section 3">Impact on the Pharmaceutical Field and Beyond</div>
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                Manifold represents a truly exciting advance in the field of synthetic biology. By channeling metabolic flux through BMCs and DNA scaffolds, Manifold fixes and confines metabolic pathways in space, allowing for dramatic increases in flux through the pathway. By localizing enzymes and substrates together within BMCs, required interactions happen more frequently. Additionally, by compartmentalizing the pathway within BMCs, reactions and intermediates are isolated from the rest of the cell which is both convenient for the researchers and the microbe. It’s difficult to understate the impact Manifold will have on the field of synthetic biology and pharmacology, but we do foresee these industries as having the greatest use of Manifold. If successful, it will revolutionize how bacteria are engineered to synthesize anything from bulk chemicals to pharmaceuticals to biofuels. The implications on the pharmaceutical world alone are astounding. The cost of production of virtually any drug synthesized in bacteria can be reduced, in turn, reducing the cost of life-saving as well as everyday drugs. While creating our proof of concept design for resveratrol, we focused mainly on the pharmaceutical industry, determining how the synthesis of this supplement will impact the surrounding field, but if the use of Manifold proves successful, any form of enzymatic compound production has the potential to be heightened. Through the combination of BMCs and DNA scaffolds to channel metabolic flux, Manifold represents an exciting advance in the field that can not be underestimated. Though there is much work to be done yet, the team is thrilled to continue crafting its approach to Manifold.
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              <b>Fig 1.</b> An example of a bioreactor in which bacteria produced through the Manifold system would be grown in order to produce pharmaceuticals.
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          <div class="sectionTitle" id="Section 4">Safety Considerations</div>
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              There arise a number of safety and ethical considerations that will require close attention. As pointed out before, we have questioned who will first benefit from our technology? Are there ways in which Manifold can be manipulated for malicious purposes? In looking at potential malpractice by an entity licensing our platform, we see little risk in negligent or even intentional release of our bacteria into the wild. Fortunately, with the high metabolic load placed on our engineered <i>E. coli</i> cells, it follows that our organism would face a significant disadvantage if released into the environment. For this reason, there arises no reason to suspect that the engineered organism is any cause for concern. Unfortunately, there exist perhaps an equal number of ways to commandeer the technology for selfish or malicious motivations as there are ways to use the technology for well-meaning adaptations. Here lies the worst case scenario regarding Manifold. In the wrong hands, this technology can be used to increase production of compounds to be used as weapons. Through Manifold, dangerous compounds that stem from enzymatic pathways can be produced far more efficiently, and as creators of this platform, we do feel a responsibility to manage the ways in which our technology is used. This sentiment became our Code of Ethical Conduct, our internal doctrine that governs our own use of the platform, while also giving guidelines about judging what outside groups can utilize Manifold. Foundational advances are powerful tools; they derive much of their impact and excitement from the possibility to adapt the advance to an array of purposes, organisms, and disciplines.
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<h1>Proposed Implementation</h1>
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<h3>Silver Medal Criterion #4</h3>
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<p>Explain how you would implement your project in the real world.
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<br><br>
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Please see the <a href="https://2020.igem.org/Judging/Medals">2020 Medals Page</a> for more information. </p>
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Latest revision as of 03:31, 28 October 2020

Manifold

Index:
Implementation
Testing Efficacy
Beyond a proof of concept to determine the success of Manifold, further tests will be required to determine the effectiveness of our design and its scalability. The primary test for efficacy will be an HPLC analysis of the culture broth in order to identify the concentration of resveratrol being produced. To see if the Manifold system adequately increases the flux through the pathway, the final ratios of resveratrol to 4-coumaric acid can be compared. If there is no flux leakage, then this ratio will be equal to 1. In order to see if an increase in overall product is observed, the ratio of resveratrol concentrations between the experimental group and control groups can be used. Finally, to see if the rate of resveratrol is increased by the system, a linear regression can be fit to each concentration vs time plot for resveratrol and the slopes can be compared. If our yields prove that Manifold nearly eliminates flux leakage, we have the data required to move on to testing other enzyme pathways.
Further Validating Manifold
If our implementation for Manifold proves successful with resveratrol, we can move onto testing other enzyme pathways in similar fashions. Proving Manifold with pathways that are applicable to more important pharmaceutical pathways will be the priority, and we have explored options relating to statins, medications meant to reduce cholesterol levels, in order to extend our system to the production of more diverse compounds. Doing so will demonstrate the scalability of Manifold, and show that our platform technology in the space of biologics has the potential to increase yields and bolster efficiency.
Impact on the Pharmaceutical Field and Beyond
Manifold represents a truly exciting advance in the field of synthetic biology. By channeling metabolic flux through BMCs and DNA scaffolds, Manifold fixes and confines metabolic pathways in space, allowing for dramatic increases in flux through the pathway. By localizing enzymes and substrates together within BMCs, required interactions happen more frequently. Additionally, by compartmentalizing the pathway within BMCs, reactions and intermediates are isolated from the rest of the cell which is both convenient for the researchers and the microbe. It’s difficult to understate the impact Manifold will have on the field of synthetic biology and pharmacology, but we do foresee these industries as having the greatest use of Manifold. If successful, it will revolutionize how bacteria are engineered to synthesize anything from bulk chemicals to pharmaceuticals to biofuels. The implications on the pharmaceutical world alone are astounding. The cost of production of virtually any drug synthesized in bacteria can be reduced, in turn, reducing the cost of life-saving as well as everyday drugs. While creating our proof of concept design for resveratrol, we focused mainly on the pharmaceutical industry, determining how the synthesis of this supplement will impact the surrounding field, but if the use of Manifold proves successful, any form of enzymatic compound production has the potential to be heightened. Through the combination of BMCs and DNA scaffolds to channel metabolic flux, Manifold represents an exciting advance in the field that can not be underestimated. Though there is much work to be done yet, the team is thrilled to continue crafting its approach to Manifold.
Fig 1. An example of a bioreactor in which bacteria produced through the Manifold system would be grown in order to produce pharmaceuticals.
Safety Considerations
There arise a number of safety and ethical considerations that will require close attention. As pointed out before, we have questioned who will first benefit from our technology? Are there ways in which Manifold can be manipulated for malicious purposes? In looking at potential malpractice by an entity licensing our platform, we see little risk in negligent or even intentional release of our bacteria into the wild. Fortunately, with the high metabolic load placed on our engineered E. coli cells, it follows that our organism would face a significant disadvantage if released into the environment. For this reason, there arises no reason to suspect that the engineered organism is any cause for concern. Unfortunately, there exist perhaps an equal number of ways to commandeer the technology for selfish or malicious motivations as there are ways to use the technology for well-meaning adaptations. Here lies the worst case scenario regarding Manifold. In the wrong hands, this technology can be used to increase production of compounds to be used as weapons. Through Manifold, dangerous compounds that stem from enzymatic pathways can be produced far more efficiently, and as creators of this platform, we do feel a responsibility to manage the ways in which our technology is used. This sentiment became our Code of Ethical Conduct, our internal doctrine that governs our own use of the platform, while also giving guidelines about judging what outside groups can utilize Manifold. Foundational advances are powerful tools; they derive much of their impact and excitement from the possibility to adapt the advance to an array of purposes, organisms, and disciplines.