Difference between revisions of "Team:Technion Israel/S.Tar intro"

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The S.Tar platform offers the means to decide what substance will trigger a chemotactic  
 
The S.Tar platform offers the means to decide what substance will trigger a chemotactic  
 
response- controlled chemotaxis. The base of our project and the source of our name is the Tar chemoreceptor,  
 
response- controlled chemotaxis. The base of our project and the source of our name is the Tar chemoreceptor,  
one of four E. coli receptor. By changing Tar’s ligand binding domain to that of other  
+
one of four E. coli receptors. By changing Tar’s ligand binding domain to that of other  
 
receptors or by mutating it we show that E. coli can be engineered to respond to completely  
 
receptors or by mutating it we show that E. coli can be engineered to respond to completely  
 
new materials.
 
new materials.
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<p class="text-justify">
 
<p class="text-justify">
 
As a proof of concept we present two receptors:<br>
 
As a proof of concept we present two receptors:<br>
<b>1.</b> The <a href="https://2016.igem.org/Team:Technion_Israel/Modifications/pcta" target="_blank"><b>PctA-Tar</b></a>,  
+
<b>1. </b> <a href="https://2016.igem.org/Team:Technion_Israel/Proof" target="_blank"><b>PctA-Tar</b></a>,  
 
a chimera created by replacing the Tar LBD with that of the Pseudomonas receptor PctA.<br>
 
a chimera created by replacing the Tar LBD with that of the Pseudomonas receptor PctA.<br>
<b>2.</b> The <a href="https://2016.igem.org/Team:Technion_Israel/Modifications/Rosetta" target="_blank"><b>His-Tar</b></a>,  
+
<b>2.</b> <a href="https://2016.igem.org/Team:Technion_Israel/Modifications/Rosetta" target="_blank"><b>Histamine-Tar</b></a>,  
a receptor created with the help of computational biology - using the Rosetta bioinformatics  
+
a receptor created with the help of computational design - using 'Rosetta' bioinformatics  
 
software suite to design mutations in the Tar receptor.
 
software suite to design mutations in the Tar receptor.
 
</p>
 
</p>
 
</div>
 
</div>
 
</div>
 
</div>
 
<br>
 
  
 
<!-- 12 text div -->
 
<!-- 12 text div -->
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Our attempts did not end here, we also present two bold attempts to fuse a ligand binding  
 
Our attempts did not end here, we also present two bold attempts to fuse a ligand binding  
 
domain of human origin to Tar in order to induce chemotaxis towards  
 
domain of human origin to Tar in order to induce chemotaxis towards  
<a href="https://2016.igem.org/Team:Technion_Israel/Modifications/EstoTar" target="_blank"><b>estrogen</b></a>  
+
<a href="https://2016.igem.org/Team:Technion_Israel/Modifications/EstoTar" target="_blank"><b>Estrogen hormone</b></a>  
and a novel lock and key system which sadly we could not finish in time - this system relies on
+
and a novel lock and key system relies on
 
<a href="https://2016.igem.org/Team:Technion_Israel/Modifications/Intein" target="_blank"><b>intein proteins</b></a>  
 
<a href="https://2016.igem.org/Team:Technion_Israel/Modifications/Intein" target="_blank"><b>intein proteins</b></a>  
and is supposed to disable chemotaxis until a specific substance “opens the door” - induces  
+
and is purpose is to disable chemotaxis until a specific substance “opens the door” - induces  
 
intein splicing and enables chemotaxis.<br>
 
intein splicing and enables chemotaxis.<br>
 
<br>
 
<br>

Revision as of 09:04, 16 October 2016

S.tar, by iGEM Technion 2016

S.tar, by iGEM Technion 2016

Chemotaxis is the movement of an organism in response to an external chemical stimulus The bacterial chemotaxis system is characterized by specific responses, high sensitivity and a dynamic range (1). This system enables bacteria to sense their immediate environment and quickly adapt to changes in its chemical composition - fleeing from repellents and moving towards attractants. The key to this system and the first link in the chain is the bacterial chemoreceptor - a membrane protein that binds the effector and transduces the signal down the line.
Despite its advantages this system is still rather limited, bacterial chemoreceptors have evolved to sense specific materials that either benefit or harm the cell in some way. The specificity of a receptor is determined by its ligand binding domain. This domain is the focus of project S.Tar.

Fig. 1: A bacterial sense of direction.



S.Tar

The S.Tar platform offers the means to decide what substance will trigger a chemotactic response- controlled chemotaxis. The base of our project and the source of our name is the Tar chemoreceptor, one of four E. coli receptors. By changing Tar’s ligand binding domain to that of other receptors or by mutating it we show that E. coli can be engineered to respond to completely new materials.

As a proof of concept we present two receptors:
1. PctA-Tar, a chimera created by replacing the Tar LBD with that of the Pseudomonas receptor PctA.
2. Histamine-Tar, a receptor created with the help of computational design - using 'Rosetta' bioinformatics software suite to design mutations in the Tar receptor.

Our attempts did not end here, we also present two bold attempts to fuse a ligand binding domain of human origin to Tar in order to induce chemotaxis towards Estrogen hormone and a novel lock and key system relies on intein proteins and is purpose is to disable chemotaxis until a specific substance “opens the door” - induces intein splicing and enables chemotaxis.

We have created non-natural chemoreceptors, by modifying Tar’s ligand binding domain.




The main goal of project S.Tar was to design new chemoreceptors that do not exist in nature. These new chemoreceptors were based on the Tar chemoreceptor.
Said chemoreceptor structure consists of three main regions: Ligand binding domain (LBD), The HAMP domain and the MH bundle (2).

Fig. 3: Tar chemoreceptor structure.




Our project focused on altering the LBD region by using different tools and approaches each designed to achieve a unique response. The three major approaches were as follows:
1. Tar-Chimeras: a hybrid receptor consisting of an LBD from different sources, such as human and other bacteria, fused to the HAMP and MH of Tar. An example of these hybrids: PctA-Tar and Narx-Tar.
2. Intein: A protein fused into the LBD that functions as an on/off mechanism that allows the control of chemotaxis.
3. Rosetta: a computer software that helps with the design of new LBDs that recognizes the ligand of your choice.

Referances

1. Bi, S. and Lai, L., 2015. Bacterial chemoreceptors and chemoeffectors. Cellular and Molecular Life Sciences, 72(4), pp.691-708.

2. BI, Shuangyu; LAI, Luhua. Bacterial chemoreceptors and chemoeffectors. Cellular and Molecular Life Sciences, 2015, 72.4: 691-708.‏

3. British Society for Immunology




S.tar, by iGEM Technion 2016