Team:TecMonterrey

From 2013.igem.org

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<h2><b>Project description</b></h2>
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It has been shown that certain bacteria such as Escherichia coli, Salmonella, and Clostridium selectively colonize and grow in tumors and a number of reports have demonstrated that some bacteria are capable of targeting both primary tumors and metastases.
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By exploiting this inherent ability of E. coli, this project aims to combine 3 different approaches in one bacterial system: the development of an E. coli capable of producing two cancer-specific apoptosis inducing proteins, a secretion system for the delivery of therapeutic proteins, and the design of a control mechanism confining the production of the therapeutic proteins only in tumor-specific regions.
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|[[Image:TecMonterrey_team.png|right|frame|iGEM TecMonterrey Team]]
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|align="center"|[[Team:TecMonterrey | Team TecMonterrey]]
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<!--- The Mission, Experiments --->
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        /*Formato del Index*/
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!align="center"|[[Team:TecMonterrey|Home]]
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/*Formato del Index*/
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!align="center"|[[Team:TecMonterrey/Team|Team]]
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!align="center"|[https://igem.org/Team.cgi?year=2013&team_name=TecMonterrey Official Team Profile]
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!align="center"|[[Team:TecMonterrey/Project|Project]]
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!align="center"|[[Team:TecMonterrey/Parts|Parts Submitted to the Registry]]
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!align="center"|[[Team:TecMonterrey/Modeling|Modeling]]
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!align="center"|[[Team:TecMonterrey/Notebook|Notebook]]
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!align="center"|[[Team:TecMonterrey/Safety|Safety]]
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                  <a class="tag" href="#">Home</a>
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                  <div class="menuhover">Project</div>
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                  <a class="tag" href="./Team:TecMonterrey/Project.html">Project</a>
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                  <ul class="sub_navigation">
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                    <li class="list"><a href="./Team:TecMonterrey/Project.html#BKG">Background</a></li>
 +
                    <li class="list"><a href="./Team:TecMonterrey/Project.html#DSC">Description</a></li>
 +
                    <li class="list"><a href="./Team:TecMonterrey/Project.html#MDLL">Modelling</a></li>
 +
                      <li class="list"><a href="./Team:TecMonterrey/Project.html#STY">Security and Safety</a></li>
 +
                      <li class="list"><a href="./Team:TecMonterrey/Project.html#OWN">Novel ownership and sharing approach</a></li>
 +
                      <li class="list"><a href="./Team:TecMonterrey/Project.html#FW">Future Work</a></li>
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                  </ul>
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              </li>
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 +
                    <li class="list"><a href="./Team:TecMonterrey/methods.html#GNL">General Methods</a></li>
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 +
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 +
                      <li class="list"><a href="./Team:TecMonterrey/methods.html#INTL">Inernalization</a></li>
 +
                  </ul>
 +
              </li>
 +
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 +
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 +
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 +
                  </ul>
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 +
                    <li class="list"><a href="./Team:TecMonterrey/human_practice.html#EC">Cancer Manual</a></li>
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                  </ul>
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 +
        </div>
 +
    </div>
 +
    <div id="dude" style="margin-bottom:40px;">
 +
        <h2>Modular, synthetic biology approach for the development of a bacterial cancer therapy in <i>Escherichia coli.</i></h2>
 +
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                <p class="lbl" style="top:115px; left:225px;">IV</p>
 +
                <p class="lbl" style="top:210px; left:80px;">I</p>
 +
                <p class="lbl" style="top:230px; left:240px;">III</p>
 +
                <p class="lbl" style="top:305px; left:295px;">II</p>
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            </div>
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            <div id="text">
 +
                <h3 style="text-align:center;">Overview</h3>
 +
                <p style="font-size:12px;">By harnessing the inherent ability of facultative anaerobic bacteria to colonize and grow in tumoral environments, this project aims to prove the functionality of four different modules that would work together as a bacterial cancer therapy using Escherichia coli as chasis: Toxicity module, Secretion module, Localized induction module, and Internalization module.
 +
The expression of tumor specific therapeutic proteins, Apoptin and TRAIL, conforms the toxicity module. For these proteins to have their effect they need to be located in the extracellular matrix, therefore we are developing a module with a secretion function using hemolysin secretory mechanism. The hypoxic microenvironment present in tumors can be used for the localized induction module of tumor specific proteins, using the promoters HIP and nirB. Finally, Apoptin needs mechanisms to enter tumor cells’ cytoplasm. Proteins with this requirement could reach the cytoplasm when coupled with the internalization module, resulting in a fusion with the TAT peptide.</p>
 +
            </div>
 +
    </div>
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 +
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        <li style="float:left;list-style:none;"><img src="https://static.igem.org/mediawiki/2013/a/a9/Promega1.jpg" style="width:120px;height:60px;margin-top:20px;"></li>
 +
        <li style="float:left;list-style:none;"><img src="https://static.igem.org/mediawiki/2013/3/3d/GenScript.png" style="width:140px;height:60px;margin-top:20px;"></li>
 +
        <li style="float:left;list-style:none;"><img src="https://static.igem.org/mediawiki/2013/9/96/Catedra.png" style="width:180px;height:60px;margin-top:20px;"></li>
 +
        <li style="float:left;list-style:none;"><img src="https://static.igem.org/mediawiki/2013/6/61/Logo_NL_Logramos_Mas_Baja.png" style="width:140px;height:60px;margin-top:20px;"></li>
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    </ul></div>
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                "<h3 style=\"text-align:center;\">Overview</h3><p style=\"font-size:12px;\">By harnessing the inherent ability of facultative anaerobic bacteria to colonize and grow in tumoral environments, this project aims to prove the functionality of four different modules that would work together as a bacterial cancer therapy using <i>Escherichia coli</i> as chasis: Toxicity module, Secretion module, Localized induction module, and Internalization module. The expression of tumor specific therapeutic proteins, Apoptin and TRAIL, conforms the toxicity module. For these proteins to have their effect they need to be located in the extracellular matrix, therefore we are developing a module with a secretion function using hemolysin secretory mechanism. The hypoxic microenvironment present in tumors can be used for the localized induction module of tumor specific proteins, using the promoters HIP and nirB. Finally, Apoptin needs mechanisms to enter tumor cells’ cytoplasm. Proteins with this requirement could reach the cytoplasm when coupled with the internalization module, resulting in a fusion with the TAT peptide.</p>",
 +
"<h3 style=\"text-align:center;\">Tumor Specific Induction</h3><p style=\"font-size:16px;\">An important part of this Project is the localized expression of the therapeutic proteins. For more than sixty years it has been known that some bacteria grow preferably in hypoxic conditions, being <i>E. coli</i> one of these organisms. Tumors are known to be regions with low oxygen concentrations (hypoxic). With these factors in favor, three different hypoxia promoters were characterized.</p>",
 +
"<h3 style=\"text-align:center;\">Therapeutic Proteins Production</h3><p style=\"font-size:16px;\">The bacterial system expresses two different proteins with the help of a hypoxia promoter. This experiment aims to prove the expression of the two toxins in <i>E. coli</i> (Apoptin and TRAIL). The expression of these tumor specific therapeutic proteins conforms the “Therapeutic proteins” module. More than 70 human tumor cell lines were shown to be susceptible to Apoptin, whereas Apoptin does not induce apoptosis in a variety of normal cells. TRAIL is a tumor specific agent under development as a novel anticancer therapeutic agent.</p>",
 +
"<h3 style=\"text-align:center;\">Enhanced Secretion System</h3><p style=\"font-size:16px;\">In order for TRAIL and Apoptin to exert their anticancer activity they must reach the extracellular matrix in the first place. This module uses the type I alpha-hemolysin secretion system of <i>E. coli</i> to achieve that feat.</p>",
 +
"<h3 style=\"text-align:center;\">Internalization</h3><p style=\"font-size:16px;\">Internalization of Apoptin into the cytosol of human cells was confirmed using a Tat-GFP proof of concept approach.<br><br>The internalization module was characterized trough the use of Tat-GFP fusion protein puriifed by His-tag (His Pur, Thermo Scientific). Internalization assay was performed on coverslip grown human cells (NIH3, Caco-2) treated with 1&#181;g/mL,5&#181;g/mL and 10&#181;g/mL Tat-GFp.</p>"
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Latest revision as of 03:25, 29 October 2013

iGem mty

Modular, synthetic biology approach for the development of a bacterial cancer therapy in Escherichia coli.

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Overview

By harnessing the inherent ability of facultative anaerobic bacteria to colonize and grow in tumoral environments, this project aims to prove the functionality of four different modules that would work together as a bacterial cancer therapy using Escherichia coli as chasis: Toxicity module, Secretion module, Localized induction module, and Internalization module. The expression of tumor specific therapeutic proteins, Apoptin and TRAIL, conforms the toxicity module. For these proteins to have their effect they need to be located in the extracellular matrix, therefore we are developing a module with a secretion function using hemolysin secretory mechanism. The hypoxic microenvironment present in tumors can be used for the localized induction module of tumor specific proteins, using the promoters HIP and nirB. Finally, Apoptin needs mechanisms to enter tumor cells’ cytoplasm. Proteins with this requirement could reach the cytoplasm when coupled with the internalization module, resulting in a fusion with the TAT peptide.