Difference between revisions of "Team:SPSingapore/Description"

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<h2> Project Description </h2>
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<p>Tell us about your project, describe what moves you and why this is something important for your team.</p>
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<ul>
 
<ul>
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<li> A clear and concise description of your project.</li>
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<li class = 'active'><a href='https://2015.igem.org/Team:SPSingapore/'><span>Home</span></a></li>
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<li>A detailed explanation of why your team chose to work on this particular project.</li>
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<li><a href='https://2015.igem.org/Team:SPSingapore/Team'><span>Team</span></a>
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<li>References and sources to document your research.</li>
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<ul>
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<li>Use illustrations and other visual resources to explain your project.</li>
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<li><a href="https://2015.igem.org/Team:SPSingapore/Team">Overview</a></li>
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<li><a href="https://igem.org/Team.cgi?id=1804">Official Profile</a></li>
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        <li><a href="https://2015.igem.org/Team:SPSingapore/Team Bios">Team Bios</a></li>
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        <li><a href="https://2015.igem.org/Team:SPSingapore/Mentors">Mentors</a></li>
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        <li><a href="https://2015.igem.org/Team:SPSingapore/Attributions">Attributions</a></li>
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</li>
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<li><a href='https://2015.igem.org/Team:SPSingapore/Project'><span>Project</span></a>
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<ul>
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<li><a href='https://2015.igem.org/Team:SPSingapore/Project'>Overview</a></li>
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        <li><a href="https://2015.igem.org/Team:SPSingapore/Invasin">Invasin + Listerolysin</a></li>
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        <li><a href="https://2015.igem.org/Team:SPSingapore/ESAQS">esa Quorum Sensing</a></li>
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        <li><a href="https://2015.igem.org/Team:SPSingapore/Anaerobic Promoter">Anaerobic Promoter</a></li>
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        <li><a href="https://2015.igem.org/Team:SPSingapore/Parts">Parts</a></li>
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</ul>
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</li>
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<li><a href='https://2015.igem.org/Team:SPSingapore/Notebook'><span>Notebook</span></a>
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<ul>
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<li><a href="https://2015.igem.org/Team:SPSingapore/Protocol">Protocols</a></li>
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        <li><a href="https://2015.igem.org/Team:SPSingapore/Notebook">Entries</a></li>
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</ul>
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</li>
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<li><a href='https://2015.igem.org/Team:SPSingapore/Practices'><span>Human Practices</span></a>
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<ul>
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<li><a href="https://2015.igem.org/Team:SPSingapore/Practices">Overview</a></li>
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<li><a href="https://2015.igem.org/Team:SPSingapore/Workshop">Workshop</a></li>
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<li><a href="https://2015.igem.org/Team:SPSingapore/Workshop Materials">Workshop Materials</a></li>
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        <li><a href="https://2015.igem.org/Team:SPSingapore/Interview">Consultations</a></li>
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</ul>
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</li>
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<li><a href='https://2015.igem.org/Team:SPSingapore/Safety'><span>Safety</span></a></li>
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<li class='last'><a href='https://2015.igem.org/Team:SPSingapore/Medals'><span>Medals</span></a></li>
 
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  <li><a href = "https://2015.igem.org/Team:SPSingapore/Practices"><span>HUMAN PRACTICES</span></a>
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        <li><a href = "https://2015.igem.org/Team:SPSingapore/Workshop"><span>Workshop </span></a></li>
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        <li><a href = "https://2015.igem.org/Team:SPSingapore/Workshop Materials"><span>Workshop Materials</span></a></li>
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        <li class = 'last'><a href = "https://2015.igem.org/Team:SPSingapore/Interview"><span>Consultations</span></a></li>
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      </ul>
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  </li>
  
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<li class='last'><a><span style = "background-color:midnightblue"><br>"An expert is someone who knows some of the worst mistakes that can be made in his subject and how to avoid them.<br><br> - Werner Heisenberg</span></a>
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<h4>Advice on writing your Project Description</h4>
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<td colspan=1 style = "box-shadow: 0 0 0; padding:0;border-top:5px white;font-size:15px"><h1>Consultations with Experts</h1></td>
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The Invasin-Listerolysin system
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The Invasin protein from Yersinia is a outer surface protein that induces phagocytosis of the pathogenic Yersinia by mammalian cells in a beta integrin dependent manner(<a href = "#ref">Wiedemann et al., 2001</a>). Whereas the Listerolysin O cytolysin of Listeria moncytogenes is a well documented cytolysin, which allows for lysis of the endosome vacuole by the pathogen after entry into a mammalian cell (<a href = "#ref">Dramsi & Cossart, 2002</a>). This facilitates endosomal escape of the bacteria. When these two [proteins are coupled together, they can confer non-pathogenic bacteria with suitable features to serve as a drug delivery vector to the cytoplasm.
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Registry Update of BBa_299812
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In constructing our Anaerobic Response System, we chose Invasin and Listeriolysin as part of our drug delivery system, building upon a construct from the Warsaw 2010 team, BBa_K299812. BBa_K299812 is a useful intermediate part containing the Invasin and Listeriolysin with ribosome binding sites in front of the genes. Different promoter sequences could be prepended ahead of the part and modified for our use.
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<br><br>
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However, when sequencing the part we received from the repository, our team found that the part sequence had major mismatches with the documented sequence. This was particularly serious for the invasin gene, which had only 55% identity between the translated amino acid sequences from the part wiki and our sequencing results. Furthermore, even though BLAST results showed that the invasin gene was indeed present, the strain of origin seemed to be different from the documentation. Lastly, the part does not strictly adhere to BioBricks standard, because the suffix sequence had only the PstI site left. (Further details and updated sequence can be found <a href = "http://parts.igem.org/Part:BBa_K299812:Experience">here</a>.)
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<br><br>
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As our lab only has the resources to functionally characterise the effect of the expression of invasin through an invasion assay, invasin was our main focus for the construction of parts for our Anaerobic Response System. 
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References
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Dramsi, S. and Cossart, P. (2002) Listeriolysin O: a genuine cytolysin optimized for an intracellular parasite. J Cell Biol. 2002 Mar 18; 156(6): 943–946.
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<br>Wiedemann A1, Linder S, Grassl G, Albert M, Autenrieth I, Aepfelbacher M. (2001) Yersinia enterocolitica invasin triggers phagocytosis via beta1 integrins, CDC42Hs and WASp in macrophages. Cell Microbiol. 2001 Oct;3(10):693-702.
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Future Work
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We encourage you to put up a lot of information and content on your wiki, but we also encourage you to include summaries as much as possible. If you think of the sections in your project description as the sections in a publication, you should try to be consist, accurate and unambiguous in your achievements.  
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<img src = "https://static.igem.org/mediawiki/2015/a/ab/SPSingapore_Team-Logo-New.png" height = "60px" style="margin-top:5px;margin-left:-130px;">
 
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Judges like to read your wiki and know exactly what you have achieved. This is how you should think about these sections; from the point of view of the judge evaluating you at the end of the year.
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<img src="https://static.igem.org/mediawiki/2015/0/05/SPSingapore_NUS-Logo.png" height="50px" style="margin-top:10px;margin-right:30px;padding-right:30px;border-right: 3px solid lightgrey;">
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<img src="https://static.igem.org/mediawiki/2015/7/72/SPSingapore_SPS-Logo.png" height="50px" style="margin-top:10px;margin-right:30px;">
 
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<h4>References</h4>
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<p>iGEM teams are encouraged to record references you use during the course of your research. They should be posted somewhere on your wiki so that judges and other visitors can see how you though about your project and what works inspired you.</p>
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<h4>Inspiration</h4>
 
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<p>See how other teams have described and presented their projects: </p>
 
  
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<li><a href="https://2014.igem.org/Team:Imperial/Project"> Imperial</a></li>
 
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<li><a href="https://2014.igem.org/Team:UC_Davis/Project_Overview"> UC Davis</a></li>
 
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<li><a href="https://2014.igem.org/Team:SYSU-Software/Overview">SYSU Software</a></li>
 
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Revision as of 00:57, 19 September 2015


Consultations with Experts


The Invasin-Listerolysin system
The Invasin protein from Yersinia is a outer surface protein that induces phagocytosis of the pathogenic Yersinia by mammalian cells in a beta integrin dependent manner(Wiedemann et al., 2001). Whereas the Listerolysin O cytolysin of Listeria moncytogenes is a well documented cytolysin, which allows for lysis of the endosome vacuole by the pathogen after entry into a mammalian cell (Dramsi & Cossart, 2002). This facilitates endosomal escape of the bacteria. When these two [proteins are coupled together, they can confer non-pathogenic bacteria with suitable features to serve as a drug delivery vector to the cytoplasm.

Registry Update of BBa_299812
In constructing our Anaerobic Response System, we chose Invasin and Listeriolysin as part of our drug delivery system, building upon a construct from the Warsaw 2010 team, BBa_K299812. BBa_K299812 is a useful intermediate part containing the Invasin and Listeriolysin with ribosome binding sites in front of the genes. Different promoter sequences could be prepended ahead of the part and modified for our use.

However, when sequencing the part we received from the repository, our team found that the part sequence had major mismatches with the documented sequence. This was particularly serious for the invasin gene, which had only 55% identity between the translated amino acid sequences from the part wiki and our sequencing results. Furthermore, even though BLAST results showed that the invasin gene was indeed present, the strain of origin seemed to be different from the documentation. Lastly, the part does not strictly adhere to BioBricks standard, because the suffix sequence had only the PstI site left. (Further details and updated sequence can be found here.)

As our lab only has the resources to functionally characterise the effect of the expression of invasin through an invasion assay, invasin was our main focus for the construction of parts for our Anaerobic Response System.

References
Dramsi, S. and Cossart, P. (2002) Listeriolysin O: a genuine cytolysin optimized for an intracellular parasite. J Cell Biol. 2002 Mar 18; 156(6): 943–946.
Wiedemann A1, Linder S, Grassl G, Albert M, Autenrieth I, Aepfelbacher M. (2001) Yersinia enterocolitica invasin triggers phagocytosis via beta1 integrins, CDC42Hs and WASp in macrophages. Cell Microbiol. 2001 Oct;3(10):693-702.

Future Work