From cb072e2f93b496fd36ee52ad5c509b38f14b119a Mon Sep 17 00:00:00 2001
From: Bindert Algra <bindert_algra@outlook.com>
Date: Fri, 14 Oct 2022 02:11:54 +0000
Subject: [PATCH] Update wiki/pages/partnership.html
---
wiki/pages/partnership.html | 8 ++++----
1 file changed, 4 insertions(+), 4 deletions(-)
diff --git a/wiki/pages/partnership.html b/wiki/pages/partnership.html
index 0503f18..71e1f6b 100644
--- a/wiki/pages/partnership.html
+++ b/wiki/pages/partnership.html
@@ -7,8 +7,8 @@
<div class="project_desc">
<div class="long_desc">
- <h1 style="background-color: black; margin-left: -10%; margin-right: 65%; padding-top: 2%; padding-bottom: 2%; padding-left: 12%; padding-right: 5%;"><b><u>The beginning of</u></b></h1>
- <h1 style="background-color: black; margin-top: -5%; margin-left: -10%; margin-right: 65%; padding-top: 2%; padding-bottom: 2%; padding-left: 12%;"><b><u>something beautiful</u></b></h1>
+ <h1 style="background-color: #ffc30b; margin-left: -10%; margin-right: 65%; padding-top: 2%; padding-bottom: 2%; padding-left: 12%; padding-right: 5%;"><b><u>The beginning of</u></b></h1>
+ <h1 style="background-color: #ffc30b; margin-top: -5%; margin-left: -10%; margin-right: 65%; padding-top: 2%; padding-bottom: 2%; padding-left: 12%;"><b><u>something beautiful</u></b></h1>
<p>The idea of a partnership with the Wageningen 2022 iGEM team, Colourectal, quickly arose after finding out that both projects must deal with the same challenge: confining a probiotic GMO within the body of its host. Both teams had already done extensive research regarding the topic of biocontainment and concluded to use temperature as a variable to activate our kill systems. After meeting again during the iGEM meetup in Utrecht plans were made to set up a video call (figure 1) to discuss what grew out to be a fruitful partnership as characterized by the event in the timeline below (figure 2).</p>
<img style="margin-left: 25%; padding-left: 25%; padding-right: 0%;" src="https://static.igem.wiki/teams/4233/wiki/partnership-1.png" width="50%">
@@ -17,7 +17,7 @@
<p style="font-style: italic; color: white; margin-left: 10%;">Figure 2. Timeline of the partnership with team Wageningen.</p>
</div>
<div style="margin-top: 5%;" class="long_desc">
- <h1 style="background-color: black; margin-left: -10%; margin-right: 65%; padding-top: 2%; padding-bottom: 2%; padding-left: 12%; padding-right: 5%;"><b><u>Lab Cooperation</u></b></h1>
+ <h1 style="background-color: #ffc30b; margin-left: -10%; margin-right: 65%; padding-top: 2%; padding-bottom: 2%; padding-left: 12%; padding-right: 5%;"><b><u>Lab Cooperation</u></b></h1>
<p>During this first meeting we found out that we shared the use of the same temperature sensitive promoter pTlpA, developed by Piraner, et al. (2017) [1] and converted into a biobrick (BBa_K2500003) by iGEM 2017 Zurich. However, our utilization of this sensor is quite different in principle. In our system the pTlpA(36℃, 39℃, 41℃) is used to control the expression of a TetR gene, which in turn inhibits the expression of a fatal CRISPR-Cas9 casette. The TetR gene should be repressed at temperatures lower than the chicken's body temperature which activates Cas9 and causes cell death.</p>
<p>Colourectal uses the temperature sensitive pTlpA(36℃) promoter to control the expression of the antitoxin ccdA, while the toxin ccdB is under a constitutive promoter. When the temperature drops, the promoter blocks the expression of the antitoxin ccdA, leading to cell death. Therefore, we decided to compare the two systems regarding effectiveness and leakiness.</p>
<p>We did this by both replacing the kill-mechanisms of our temperature sensors by fluorescent reporter genes, with Colourectal choosing GFP and Nanobuddy using mRFP1, the latter of which is explained in more detail in the <a href="">Engineering</a> page. </p>
@@ -33,7 +33,7 @@ Additionally, another obvious peculiarity of our data is the initially low start
</p>
<p>Finally, to allow for a more comprehensive data-comparison it is recommended to use a shared internal standard, for example using the serial dilution of the fluorescent dyes from the Interlab study. It is also possible to use a well characterized constitutive promoter for comparison, e.g. using constructs containing BBa_J23100 or BBa_J23102, which we have also constructed. Resulting in a fluorescence fold-change relative to previously mentioned standard. However due to time-constraints this could not be performed at this time.
</p>
-<h1 style="background-color: black; margin-left: -10%; margin-right: 65%; padding-top: 2%; padding-bottom: 2%; padding-left: 12%; padding-right: 5%;"><b><u>Further Shared Experiences</u></b></h1>
+<h1 style="background-color: #ffc30b; margin-left: -10%; margin-right: 65%; padding-top: 2%; padding-bottom: 2%; padding-left: 12%; padding-right: 5%;"><b><u>Further Shared Experiences</u></b></h1>
<p>Beyond individual lab-work and comparing data, we also regularly met online throughout the summer to discuss and troubleshoot the two biocontainment projects. And when the Wageningen team was not satisfied with the backbone they used for their killswitch, the proposed solution was to try our pTRKH3 plasmid that contains a broad-range host ORI. We sent the plasmid and shared our protocols and snapgene files through a shared google drive folder.</p>
<p>Finally, we also formed a collective Whatsapp group for organization, real-time discussion of various matters, and meme-sharing. We also met physically in Groningen in June, and Hamburg and Utrecht in July to socialize and share ideas. This brought our teams more closely together, and brought our iGEM experience to new heights!</p>
<p>Sonia (Wageningen): This partnership allowed us to troubleshoot the pTlpA construct and to test it with a different backbone. Unfortunately, we haven’t managed to make our construct work, making pcspA is the best option for our project. The monthly meetings and updates with NanoBuddy made us discover how important is to discuss your project and results and get an outside point of view! </p>
--
GitLab