Difference between revisions of "Team:BostonU/Description"

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<div id="backgroundimage1"><div class="background-gradient-down"><p class="wide-heading-type mainwrap align-center">PROJECT DESCRIPTION</p></div></div>
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  <p class="body-type mainwrap indented">The presence of specific RNAs in cells can be indicative of their state. Detecting these RNAs on a small scale allows for identification of viruses such as Zika and Ebola, however measuring larger sets of RNA remains complex. Current methods of RNA detection are time consuming and require expensive machinery and technical expertise. Toehold switches offer an alternative means of RNA detection. Toehold switches are de novo designed riboregulators that exhibit high specificity, wide dynamic range, and ease of use.</p>
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  <p class="body-type mainwrap">&nbsp;</p>
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  <p class="body-type mainwrap"> FIGURE</p>
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  <p class="body-type mainwrap">&nbsp;</p>
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  <p class="body-type mainwrap indented">Toehold switches are composed of two parts: the first is a strand of mRNA called a switch that forms a hairpin loop secondary structure in which the start codon for a downstream gene is sequestered. The other component is a linear section of mRNA known as a trigger that binds to the beginning of the switch, causing the switch to unfold and allowing for protein expression from the gene contained downstream of the hairpin. If the expression cassette codes for a fluorescent protein, a fluorescence readout can allow for the detection of the trigger mRNA.</p>
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  <p class="body-type mainwrap indented">Toeholds have been shown to be effective in detecting viral RNA for Zika and Ebola (Pardee et al), by measuring 10-20 RNAs. At this point, utilizing toehold switches to detect large sets of RNA is not feasible. We aim to ease this problem by developing a logic framework that allows for the detection of multiple mRNAs to drive molecular computing. In our system, we will utilize trigger-toehold pairs to drive the downstream expression of recombinase proteins. These proteins control the expression of genes flanked by recombinase-recognition sites. This results in a system that produces variable measurable responses based on the presence of specific mRNA sequences. </p>
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  <p class="body-type mainwrap indented">&nbsp;</p>
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  <p class="body-type mainwrap indented">FIGURE </p>
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  <p class="body-type mainwrap indented">&nbsp;</p>
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  <p class="body-type mainwrap indented">Furthermore, we integrate this RNA-inducible logic within a cell-free transcription translation system to reduce the experimental burden on time and supplies. This work serves to establish functionality of RNA-inducible logic in a cell-free system, and provides a platform for future implementation in applications that require detection of multiple RNAs, including disease diagnostics. </p>
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Revision as of 06:38, 16 October 2017

PROJECT DESCRIPTION

The presence of specific RNAs in cells can be indicative of their state. Detecting these RNAs on a small scale allows for identification of viruses such as Zika and Ebola, however measuring larger sets of RNA remains complex. Current methods of RNA detection are time consuming and require expensive machinery and technical expertise. Toehold switches offer an alternative means of RNA detection. Toehold switches are de novo designed riboregulators that exhibit high specificity, wide dynamic range, and ease of use.

 

FIGURE

 

Toehold switches are composed of two parts: the first is a strand of mRNA called a switch that forms a hairpin loop secondary structure in which the start codon for a downstream gene is sequestered. The other component is a linear section of mRNA known as a trigger that binds to the beginning of the switch, causing the switch to unfold and allowing for protein expression from the gene contained downstream of the hairpin. If the expression cassette codes for a fluorescent protein, a fluorescence readout can allow for the detection of the trigger mRNA.

Toeholds have been shown to be effective in detecting viral RNA for Zika and Ebola (Pardee et al), by measuring 10-20 RNAs. At this point, utilizing toehold switches to detect large sets of RNA is not feasible. We aim to ease this problem by developing a logic framework that allows for the detection of multiple mRNAs to drive molecular computing. In our system, we will utilize trigger-toehold pairs to drive the downstream expression of recombinase proteins. These proteins control the expression of genes flanked by recombinase-recognition sites. This results in a system that produces variable measurable responses based on the presence of specific mRNA sequences.

 

FIGURE

 

Furthermore, we integrate this RNA-inducible logic within a cell-free transcription translation system to reduce the experimental burden on time and supplies. This work serves to establish functionality of RNA-inducible logic in a cell-free system, and provides a platform for future implementation in applications that require detection of multiple RNAs, including disease diagnostics.