Difference between revisions of "Team:Freiburg/Description"

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<h5  style="text-align: center"> Overview </h5>
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<h5  style="text-align: center"> Biobricks </h5>
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I) Constructed BioBricks
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Our project required the construction of fusion proteins that are displayed on the surface of the spores of B. subtilis. Anchoring an epitope-specific nanobody and glutathione S-transferase to coat proteins enables the spores to attach to surfaces and activate prodrugs on site.<br>
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In addition to targeted drug delivery, site specific enzymatic activities play a crucial role in other scientific field and many further applications are imaginable ranging from the production of metabolites to bioremediation. Therefore, we would like to share our achievements with the iGEM community in the hope to support and facilitate future projects.<br>
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Our supervisors Dr. Maximilian Ulbrich and Dr. Nicole Gensch provided us plasmids encoding the anti-GFP nanobody and the glutathione S-transferase that were inevitable for our project. The Integration vectors used to insert our constructs into the genome of B. subtlis were provided by Julia Bartels from the Mascher lab at the TU Dresden. <br>
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Below you will find a table summarizing the submitted biobricks and guiding you to the entries in the registry. <br>
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II) Improved BioBricks
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<a href="https://2016.igem.org/Team:Freiburg/Improved_Biobrick_pelB_nanobody"> pelB_anti-GFP nanobody</a><br><br>
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We submitted an improved anti-GFP nanobody containing the pelB leader sequence for the export of the protein in the periplasmatic space resulting in an increased yield of the over-expressed protein.
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<a href="https://2016.igem.org/Team:Freiburg/Improved_Biobrick_PCotYZ-RBS"> PCotYZ-RBS </a><br><br>
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The B. subtilis promoter PCotYZ regulates the expression of the late stage spore coat proteins CotY and CotZ. We included a ribosome binding site which simplifies the assembly of devices and the expression of subsequent genes.
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<a href="https://2016.igem.org/Team:Freiburg/Basic_Part"> Basic Parts </a>
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<a href="https://2016.igem.org/Team:Freiburg/Composite_Part"> Composite Parts </a>
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<a href="https://2016.igem.org/Team:Freiburg/Part_Collection"> Part Collection </a>
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    <div class="para_center_20">The treatment of diseases while avoiding systemic side effects is still a major obstacle in modern medicine. After administration, conventional drugs are distributed throughout the whole body thus affecting both, diseased and healthy cells. Current strategies on targeted drug delivery are mainly based on the applications of antibody-drug conjugates or nanoparticles. However, both approaches revealed considerable challenges in their application due to short half-life and expensive production, respectively. We develop a novel platform for targeted drug delivery by implementing highly specific nanobodies directed against surface markers of affected cells. The combination with an enzymatic functionality facilitates the local activation of prodrugs, thus preventing unnecessary side effects by systemic drug dispersal. By engineering the spores of probiotic Bacillus subtilis, a member of the human microbiome, we establish a low-cost carrier for well-tolerated treatment. 
 
 
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Revision as of 17:50, 13 October 2016

Biobricks
I) Constructed BioBricks

Our project required the construction of fusion proteins that are displayed on the surface of the spores of B. subtilis. Anchoring an epitope-specific nanobody and glutathione S-transferase to coat proteins enables the spores to attach to surfaces and activate prodrugs on site.
In addition to targeted drug delivery, site specific enzymatic activities play a crucial role in other scientific field and many further applications are imaginable ranging from the production of metabolites to bioremediation. Therefore, we would like to share our achievements with the iGEM community in the hope to support and facilitate future projects.
Our supervisors Dr. Maximilian Ulbrich and Dr. Nicole Gensch provided us plasmids encoding the anti-GFP nanobody and the glutathione S-transferase that were inevitable for our project. The Integration vectors used to insert our constructs into the genome of B. subtlis were provided by Julia Bartels from the Mascher lab at the TU Dresden.
Below you will find a table summarizing the submitted biobricks and guiding you to the entries in the registry.


II) Improved BioBricks

pelB_anti-GFP nanobody

We submitted an improved anti-GFP nanobody containing the pelB leader sequence for the export of the protein in the periplasmatic space resulting in an increased yield of the over-expressed protein.

PCotYZ-RBS

The B. subtilis promoter PCotYZ regulates the expression of the late stage spore coat proteins CotY and CotZ. We included a ribosome binding site which simplifies the assembly of devices and the expression of subsequent genes.

Posted by: iGEM Freiburg

Nanocillus - 'cause spore is more!