Difference between revisions of "Team:William and Mary/Part Collection"

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                                     <p>In deciding which parts to submit to the iGEM Registry we focused on three main aspects.</p>
 
                                     <p>In deciding which parts to submit to the iGEM Registry we focused on three main aspects.</p>
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<p>First: ensuring our project is as reproducible and extensible as possible. To that end we have submitted all of new composite fluorescent protein parts that we constructed during the project. Second: Making genome integration as straightforward as possible for iGEM teams. In order to accomplish this goal we designed, tested, and validated a new integrator cassette that allows for simple genome integration using either 3A or Gibson Assembly. Third: Increasing the number of tools available for promoter-mediated regulation in synthetic biology. We created and validated an E. coli codon optimized dCas9 variant and a suite of gRNAs to target the most commonly used promoters in iGEM.   </p>
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<p>In keeping with our theme of promoter effects on transcription, we created a suite of guide RNAs for the targeted dCas9 repression of 11 of the most frequently used promoters in the Registry. For cells which express a dCas9 protein (link to the fdCas9 part page), the transformation of any of our guide RNA parts will repress the expression of any protein under the associated promoter. We also included a scrambled guide RNA to serve as a negative control for troubleshooting purposes.
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<p>(BELOW) An example of repression: fluorescence values for RFP under the R0010 promoter, in cells with and without the R0010 guide RNA”)</p>
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<p><img src = "https://static.igem.org/mediawiki/2015/c/cf/WMdCas9_Repression_Plot.png" width = 500></p>
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</p>The breadth of the variety in our part collection provides future teams with the flexibility required to actualize their network designs without inhibition from a lack of resources. </p>
 
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Revision as of 23:36, 18 September 2015

NOISE - W&M iGEM

Part Collections

In deciding which parts to submit to the iGEM Registry we focused on three main aspects.

In keeping with our theme of promoter effects on transcription, we created a suite of guide RNAs for the targeted dCas9 repression of 11 of the most frequently used promoters in the Registry. For cells which express a dCas9 protein (link to the fdCas9 part page), the transformation of any of our guide RNA parts will repress the expression of any protein under the associated promoter. We also included a scrambled guide RNA to serve as a negative control for troubleshooting purposes.

(BELOW) An example of repression: fluorescence values for RFP under the R0010 promoter, in cells with and without the R0010 guide RNA”)

The breadth of the variety in our part collection provides future teams with the flexibility required to actualize their network designs without inhibition from a lack of resources.

Integrator Suites

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Antibiotic Operons

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dCas9s

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gRNAs

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XFPs Under Various Promoters

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G^2

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