Team:BostonU/Clotho

From 2013.igem.org

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<h7><p><a href="www.eugenecad.org">Eugene</a>, a language based on Java, which is both human and machine readable. Currently the language can be used to expedite the design process for new devices by providing a list of possible transcriptional units based on specific rules and parts available.</p>
<h7><p><a href="www.eugenecad.org">Eugene</a>, a language based on Java, which is both human and machine readable. Currently the language can be used to expedite the design process for new devices by providing a list of possible transcriptional units based on specific rules and parts available.</p>
<p>The BosotnU iGEM team used Eugene to determine the number of permutations for transcriptional units that can be assembled from the MoClo parts and devices specified by the user.  To eliminate extraneous permutations, the user must specify rules pertaining to the order and relationship between parts.</p>  
<p>The BosotnU iGEM team used Eugene to determine the number of permutations for transcriptional units that can be assembled from the MoClo parts and devices specified by the user.  To eliminate extraneous permutations, the user must specify rules pertaining to the order and relationship between parts.</p>  
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<p>Eugene can be utilized via Clotho's Eugene Scriptor app. The
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<p>Eugene can be utilized via Clotho's Eugene Scriptor app.</p>
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</p></h7>
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<p>To write a Eugene script, one must first declare the parts that are being used and their properties.  Next, one must define the specific properties for each part.  To define the devices, structural rules must be made defining what parts are in each device.  Content rules describe the relationships between parts. Lastly, the user can compute permutations and print out the results.</p></h7>
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<p><center><img src="https://static.igem.org/mediawiki/2013/b/b3/Eugene1.png"></center></p>
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<p><center><img src="https://static.igem.org/mediawiki/2013/a/a4/Eugene2.png"></center></p>
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<p>We were fortunate to be able to use Eugene through Clotho's Eugene Scriptor. The first task in writing a Eugene script is to define what the parts are. Notice that the language is very straightforward and intuitive.</p>
 
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<p><center><img src="https://static.igem.org/mediawiki/2012/b/bf/1eu.png" width="500px"></center></p>
 
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<p>In the example above, we are defining our basic parts. Although the fusion/MoClo sites themselves are not basic parts, we consider them as a separate entities because they are essential when combining different MoClo parts. At this point, fusion sites as a separate part would be a more versatile method of using the language.</p>
 
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<p>After defining which parts Eugene will be combining together, the next step would be to identify the device. In this case, different basic parts are combined to form a MoClo Level 0 part.</p>
 
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<p><center><img src="https://static.igem.org/mediawiki/2012/7/7c/2eu.png" width="500px"></center></p>
 
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<p>After defining our MoClo level 0 devices, the next step is to define MoClo Level 1 devices. In this case, a Level 1 device is designated to include a promotor, RBS, gene, and terminator. Additional rules are added as needed. For instance, we are creating rules to make sure the MoClo sites of subsequent parts are the same so they are able to be ligated together. In the picture below, the rule is expressed for promoter and RBS. Additionally, we have to be consistent with the design of the MoClo parts, which means that the part can not be flanked by the same MoClo sites as shown in the second rule.</p>
 
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<p><center><img src="https://static.igem.org/mediawiki/2012/thumb/1/18/3eu.png/800px-3eu.png" width="700px"></center></p>
 
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<p><center><img src="https://static.igem.org/mediawiki/2013/3/38/Pigcode.png"></center></p>
<p><center><img src="https://static.igem.org/mediawiki/2013/3/38/Pigcode.png"></center></p>
<p><center><img src="https://static.igem.org/mediawiki/2013/f/f2/Pigexample.png"></center></p>
<p><center><img src="https://static.igem.org/mediawiki/2013/f/f2/Pigexample.png"></center></p>
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<p><h7>The <a href="http://pigeoncad.org/">PigeonCAD</a> website has many examples.</h7></p>
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<p><h7>The <a href="http://pigeoncad.org/">PigeonCAD</a> website has additional examples.</h7></p>

Revision as of 20:10, 7 September 2013



Clotho and Eugene


Clotho Introduction

Our project is focused on creating a standardized method for the characterization of genetic circuits in synthetic biology. In order to achieve this goal, it is necessary to have a platform that enables us to create the standardized data, organize it, and manage it efficiently. We have been utilizing Clotho, a synthetic biology software suite developed with tools for analyzing and organizing data.

This section aims to describe how we have been using Clotho in our project. We divided the content into the Clotho Apps we used, giving a brief explanation of how each app works and examples of situations in which they were useful for our project.

Blasto

Blasto can be used to verify DNA sequences by comparing a sequence with the expected sequence. Blasto shows the percentage of the match and the number of gaps between sequences and where those gaps occur. Users can create a collection of their parts and save sequences for easy comparison.



Eugene

Eugene, a language based on Java, which is both human and machine readable. Currently the language can be used to expedite the design process for new devices by providing a list of possible transcriptional units based on specific rules and parts available.

The BosotnU iGEM team used Eugene to determine the number of permutations for transcriptional units that can be assembled from the MoClo parts and devices specified by the user. To eliminate extraneous permutations, the user must specify rules pertaining to the order and relationship between parts.

Eugene can be utilized via Clotho's Eugene Scriptor app.

To write a Eugene script, one must first declare the parts that are being used and their properties. Next, one must define the specific properties for each part. To define the devices, structural rules must be made defining what parts are in each device. Content rules describe the relationships between parts. Lastly, the user can compute permutations and print out the results.



PigeonCAD


PigeonCAD is a very useful tool for synthetic biology because it allows the user to visualize genetic devices. Using the Synthetic Biology Open Language (SBOL) and a simple code, the user can easily build a circuit (Bhatia et al., 2013). We used Pigeon to depict our genetic circuits.

Depicted below is an example of a simple transcriptional unit depicted in SBOL with the corresponding Pigeon code.

The PigeonCAD website has additional examples.



References

[1] Bhatia, S., and Densmore, D. (2013) "Pigeon: A Design Visualizer for Synthetic Biology." ACS Synthetic Biology 2(6):348–350. doi: 10.1021/sb400024s.