Difference between revisions of "Team:Pitt/Protease/Project"
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| − | < | + | <div class="color"><h4>Project Background</h4>The second sensing system we have designed relies on transcriptional repressors. By creating a synthetic repressor that gets cleaved by a specific protease, the extract we create will be sensitive to the protease. This can be used to detect breast and colorectal cancer biomarkers such as MMP-2 and MMP-9 in patients' urine.(<a href="http://www.gynecologiconcology-online.net/article/S0090-8258(11)00584-1/abstract">Coticchia 2011</a>) This project uses several concepts to sense proteases. First of all, this system relies on a two-hybrid repressor previously described in some detail. (<a href="http://mic.sgmjournals.org/content/journal/micro/10.1099/00221287-147-6-1651#tab2">Di Lallo 2001</a>) Secondly, instead of using the two-hybrid as a way of detecting the interaction between two proteins, we created fusion proteins that contain both parts of the two-hybrid repressor. This allowed us to insert a linker sensitive to specific proteases, which would then inactivate the repressor, and allow transcription of the reporter to occur, as shown in the image below. <br/><img style="width:75%" src="https://static.igem.org/mediawiki/2015/1/14/Pitt4.png"/></div> |
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| + | <div class="color"><h4>Project State</h4>Currently, we have all the necessary plasmids in NiCo21(DE3) cells, which are ideal for this project. We have created sensor extracts using these cells; however, we have run into problems involving dead enzymes which have delayed our results. As we obtain results after the Jamboree, this website will be updated.</div> | ||
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Latest revision as of 01:22, 19 September 2015
Protease Sensor
Project Background
The second sensing system we have designed relies on transcriptional repressors. By creating a synthetic repressor that gets cleaved by a specific protease, the extract we create will be sensitive to the protease. This can be used to detect breast and colorectal cancer biomarkers such as MMP-2 and MMP-9 in patients' urine.(Coticchia 2011) This project uses several concepts to sense proteases. First of all, this system relies on a two-hybrid repressor previously described in some detail. (Di Lallo 2001) Secondly, instead of using the two-hybrid as a way of detecting the interaction between two proteins, we created fusion proteins that contain both parts of the two-hybrid repressor. This allowed us to insert a linker sensitive to specific proteases, which would then inactivate the repressor, and allow transcription of the reporter to occur, as shown in the image below.