Every year, hundreds of thousands of horseshoe crabs are caught each year and drained of up to to 30% of their blood. Why? To collect LAL, a chemical that is crucial in the detection of endotoxins in everything from drugs to medical equipment.
LAL, or Limulus Amebocyte Lysate, found in Horseshoe Crab amebocytes, is extremely sensitive to LPS, a membrane component of gram-negative bacteria. The chemical detects these harmful endotoxins by coagulating in their presence, thus creating a visible signal of bacterial contamination. The reaction proceeds as follows: First, a present endotoxin cleaves and activates a zymogen in LAL, named Factor C. Cleaved Factor C then activates Factor B, which in turn converts a proclotting enzyme in LAL into a clotting enzyme. This clotting enzyme cleaves coagulen, another protein present, and turns it into coagulin, turning the whole mixture into a coagulated signal communicating that endotoxin is present.
While this process has been relied upon for the approval of any FDA drug, the confirmation of safe pacemakers, and the sanitation of medical equipment, it is the procurement of LAL itself that has proven inefficient and dangerous of the Horseshoe Crab population.
iTesla-SoundBio is working towards creating a viable synthetic equivalent of the LAL Assay. We have chosen to focus on Factor C, a crucial part of the LAL reaction cascade, but a valuable component by itself as well. Since Factor C cleaves in the presence of endotoxin, we are working towards synthetically producing Factor C and incorporating it into a fusion protein-system that will give off a detectable signal when exposed to endotoxin.
Our project has divided itself into two phases. The first is the actual production of synthetic Factor C by bacillus subtilis. This phase has provided its own considerations, for example taking into account the large coding region, as well as inducing expression in a gram-positive system to avoid automatic cleavage of the protein itself. The second phase comprises the conception and assembly of a signal amplification system that will produce a detectable signal when Factor C is cleaved.
While there are already companies who have developed an alternative to the LAL Assay using synthetic Factor C, the high price point has encouraged us to try to produce a simpler, cheaper alternative.
Welcome to iGEM 2018!
Your team has been approved and you are ready to start the iGEM season!
Before you start
Please read the following pages:
Styling your wiki
You may style this page as you like or you can simply leave the style as it is. You can easily keep the styling and edit the content of these default wiki pages with your project information and completely fulfill the requirement to document your project.
While you may not win Best Wiki with this styling, your team is still eligible for all other awards. This default wiki meets the requirements, it improves navigability and ease of use for visitors, and you should not feel it is necessary to style beyond what has been provided.
Uploading pictures and files
You must upload any pictures and files to the iGEM 2018 server. Remember to keep all your pictures and files within your team's namespace or at least include your team's name in the file name.
When you upload, set the "Destination Filename" to T--YourOfficialTeamName--NameOfFile.jpg. (If you don't do this, someone else might upload a different file with the same "Destination Filename", and your file would be erased!)
Wiki template information
We have created these wiki template pages to help you get started and to help you think about how your team will be evaluated. You can find a list of all the pages tied to awards here at the Pages for awards link. You must edit these pages to be evaluated for medals and awards, but ultimately the design, layout, style and all other elements of your team wiki is up to you!
Editing your wiki
On this page you can document your project, introduce your team members, document your progress and share your iGEM experience with the rest of the world!
Use WikiTools - Edit in the black menu bar to edit this page
This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started:
- State your accomplishments! Tell people what you have achieved from the start.
- Be clear about what you are doing and how you plan to do this.
- You have a global audience! Consider the different backgrounds that your users come from.
- Make sure information is easy to find; nothing should be more than 3 clicks away.
- Avoid using very small fonts and low contrast colors; information should be easy to read.
- Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the iGEM 2018 calendar
- Have lots of fun!