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− | < | + | <p> MIT: Mammalian Circuitry for Treatment of COVID-19 Cytokine Storms </p> |
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<p>Developed a computational compartmentalized cytokine network that can be modified for a variety of cytokine-targeting treatments for inflammatory diseases with ten cytokines. </p> | <p>Developed a computational compartmentalized cytokine network that can be modified for a variety of cytokine-targeting treatments for inflammatory diseases with ten cytokines. </p> | ||
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</br> | </br> | ||
− | < | + | <p> Silver: </p> |
<p>Hosted the international MMM (MIT Mammalian Meetup) with 18 registered teams.</p> | <p>Hosted the international MMM (MIT Mammalian Meetup) with 18 registered teams.</p> | ||
<p>Successfully simulated the cytokine storm system with ten cytokines. </p> | <p>Successfully simulated the cytokine storm system with ten cytokines. </p> | ||
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− | < | + | <p> Gold: </p> |
<p> Engineering success integrated by reiterating circuit design through discussions with Weiss Lab. </p> | <p> Engineering success integrated by reiterating circuit design through discussions with Weiss Lab. </p> | ||
<p> Integrated human practices through discussions of implementation with the biological engineering community including bioethicists. </p> | <p> Integrated human practices through discussions of implementation with the biological engineering community including bioethicists. </p> |
Revision as of 17:16, 5 November 2020
MIT: Mammalian Circuitry for Treatment of COVID-19 Cytokine Storms
Bronze:
Developed a computational compartmentalized cytokine network that can be modified for a variety of cytokine-targeting treatments for inflammatory diseases with ten cytokines.
Characterized 15 parts associated with the PERSIST system from the Weiss Lab (Bba_K3621000-15) which can be used by future teams in mammalian circuits.
Silver:
Hosted the international MMM (MIT Mammalian Meetup) with 18 registered teams.
Successfully simulated the cytokine storm system with ten cytokines.
Contributed one review article and one original research article to the Maastricht iGEM Journal.
Contributed to the URochester Biomarker Database for characterizing cytokine storms.
Gold:
Engineering success integrated by reiterating circuit design through discussions with Weiss Lab.
Integrated human practices through discussions of implementation with the biological engineering community including bioethicists.
Successfully modeled the cytokine storm and tested different cytokine targets to identify two therapeutic targets.
To summarize the full circuit depicted in Figure 3, we will sense the cytokines IP-10 and MCP-3 which correspond to the transcription factors ELK-1 and NF-κB respectively. CasE is expressed upon activation by ELK-1, and Csy4 is expressed upon activation by NF-kB. Using the CMV promoter, Cas6 is constitutively expressed but when either or both CasE and Csy4 are present, the mRNA for Cas6 will be cut upstream of the gene sequence, so Cas6 expression will decrease. For our output fusion protein, we have another plasmid with the constitutive promoter CMV, and likewise there is an ERN cut site upstream of the gene sequence. Thus, when both IP-10 and MCP-3 are present, scFv will be produced, inhibiting our cytokine of interest.
We developed an AND gate with five PERSIST-based plasmids. IP-10 and MCP-3 signal downstream phosphorylation pathways to activate nuclear transcription factors (nTFs) ELK-1 and NF-κB, respectively. Two minimal constitutive promoters (here, CytoMegaloVirus, CMV) are then induced by nTF-responsive element binding, upstream of ERN genes. In choosing our ERN components for translation, we evaluated the sensitivity with the relative abundance of the biomarker (nTFs ELK-1 and NF-κB) in plasma. In CCS, MCP-3 has been observed about 103 fold lower than IP-10; Csy4, an experimentally stronger “OFF” ERN, was thus placed downstream of the NF-KB-responsive promoter, while CasE, a slightly weaker ERN, was used for the IP-10 sginaling pathway. Cas6, the weakest ERN of the Cas family evaluated, when produced by either of two CMV-constitutively-expressed transcripts, degrades the scFv mRNA transcripts and causes decreased translation of anti-inflammatory soluble scFv with a slight buffer to allow for IP-10 and MCP-3 elevation to be reached. To further enhance the system dynamics by decreasing the time required to reach steady state, a PEST degradation tag can be fused to the Cas6 sequence. Our AND gate is thus engineered to produce a dose-responsive output when there is adequate IP-10 and MCP-3 to result in the degradation of Cas6 transcripts.