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| <img src="https://static.igem.org/mediawiki/2020/2/26/T--UCopenhagen--Poster_Description.png"> | | <img src="https://static.igem.org/mediawiki/2020/2/26/T--UCopenhagen--Poster_Description.png"> |
| + | <figcaption style="font-family: Avenir, Arial, Helvetica, sans-serif; font-size: 12px; line-height: 1.25;"><b>Fig. 1. Visual representation of the usage of the CIDosis patch and the engineering and scientific approach. </b>1: The patch collecting sweat from the skin. 2: The layers in the patch. 3: The scientific receptor engineering. 4: The patch after it has changed color. 5: The image-analyzing app.</figcaption> |
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| <img style="width: 75%; align-self: center;" src="https://static.igem.org/mediawiki/2020/8/88/T--UCopenhagen--Poster_G_alpha_Design.png"> | | <img style="width: 75%; align-self: center;" src="https://static.igem.org/mediawiki/2020/8/88/T--UCopenhagen--Poster_G_alpha_Design.png"> |
| + | <figcaption style="font-family: Avenir, Arial, Helvetica, sans-serif; font-size: 12px; line-height: 1.25;"><b>Fig. 4. Advanced biosensor design.</b> |
| + | Split TEV protease complementation leading to cleavage of a TEV protease cut site in GPA1, thereby initiating gene expression through the yeast pheromone pathway.</figcaption> |
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| </div> | | </div> |
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| <b>Sensitivity Comparison</b><br> | | <b>Sensitivity Comparison</b><br> |
− | We compared the three designs of engineered signaling pathways in S. cerevisiae in terms of sensitivity through modeling with ordinary differential equations (ODEs). The models revealed the additional benefits of employing the yeast pheromone cascade in signal amplification (~7 orders of magnitude), thus rendering one of the designs as a clear candidate for the application in the biosensor (fig. 4). | + | We compared the three designs of engineered signaling pathways in S. cerevisiae in terms of sensitivity through modeling with ordinary differential equations (ODEs). The models revealed the additional benefits of employing the yeast pheromone cascade in signal amplification (~7 orders of magnitude), thus rendering one of the designs as a clear candidate for the application in the biosensor (fig. 5). |
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| <b>Impact of Adverse Effects</b> | | <b>Impact of Adverse Effects</b> |
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− | We modeled the effects of hypothetical cellular scenarios (e.g. reporter toxicity) on the pathways within the framework of stochastic differential equations (SDEs). There, we explored various expected and unexpected behaviors in the models, which suggested that specific failures of pathway components may lead to characteristic statistics of reporter concentrations (fig. 5). This tool has the potential to improve our troubleshooting in the future. | + | We modeled the effects of hypothetical cellular scenarios (e.g. reporter toxicity) on the pathways within the framework of stochastic differential equations (SDEs). There, we explored various expected and unexpected behaviors in the models, which suggested that specific failures of pathway components may lead to characteristic statistics of reporter concentrations (fig. 6). This tool has the potential to improve our troubleshooting in the future. |
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| <b>Protein Modeling</b><br> | | <b>Protein Modeling</b><br> |
| As the most applicable design required utilization of the yeast pheromone cascade, we had to engineer a novel GPA1 protein that would allow for signal transduction from our designed receptor system. Guided by several iterations of simulations with Rosetta Software Suite, we identified multiple regions suitable for inserting cleavage sites. | | As the most applicable design required utilization of the yeast pheromone cascade, we had to engineer a novel GPA1 protein that would allow for signal transduction from our designed receptor system. Guided by several iterations of simulations with Rosetta Software Suite, we identified multiple regions suitable for inserting cleavage sites. |
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− | However, the predictions suggested that the post-cleavage protein fragments did not exhibit the properties we expected (fig. 6). Based on these findings, we articulated a refined framework for engineering signal transduction in our biosensor. | + | However, the predictions suggested that the post-cleavage protein fragments did not exhibit the properties we expected (fig. 7). Based on these findings, we articulated a refined framework for engineering signal transduction in our biosensor. |
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