Difference between revisions of "Team:Kent/HP/Silver"

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<p>Our team has strived to make human practices the solid foundation of our project right from the start. We wanted to ensure we fully understood the implications of any project we chose to pursue while addressing current global problems.  
 
<p>Our team has strived to make human practices the solid foundation of our project right from the start. We wanted to ensure we fully understood the implications of any project we chose to pursue while addressing current global problems.  
 
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<span>Judd High School</span>
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<span>Why CRISPR-dCas13a (C2C2)?</span>
 
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<p>This year, three members of our team: Laulwa Al Salloum, Laurens Heling and Dan Brunkow, were able to work alongside Team Judd_UK in the InterLab study. They were able to use our lab and equipment to obtain the necessary readings for their InterLab while we mentored two of their members; Nikita Shukan and Mateo Hoare through the process, as well as aiding them in their managing of the different aspects of the study, such as: using the Gilson pipettes, using the plate reader, incubating the samples, etc. In return, they generously donated their InterLab DNA from Distribution kit plate 6 for us to transform as we were unsuccessful in transforming the DNA from our own kit into the E.coli DH5 alpha cells.
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<p><ul><li>We were introduced to CRISPR-Cas9 as a tool for targeting cellular mRNA translation through Dr. Peter Ellis, a lecturer in Molecular Biology and Reproduction. However, through research, we discovered that Cas9 is a DNA endonuclease, meaning it recognizes DNA targets and cuts them. It would need to be ‘tricked’ by molecular means. C2C2, however, or Cas13a is an RNA endonuclease, meaning it cuts the RNA rather than the DNA. There are two key differences when comparing Cas13 to CAS9: Firstly, Cas13 recognizes RNA rather than DNA. Secondly, once the target is recognized, it acts promiscuously and starts to ‘chew up’ all of the RNA around it, not only the intended target sequence. <br><img src=""></li>
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<li>We wanted to utilize this knowledge and build upon the CRISPR-Cas9 and produce a tool that would just track the mRNA, not cut it. We did not want to overcomplicate our methodology and deemed tagging the mRNA with GFP would be sufficient as we would be able to essentially track the mRNA movement to ensure its localization without damaging its integrity through cutting it.
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Being able to work alongside another iGEM team was incredibly useful in viewing the study from a different point of view as Judd_UK were experiencing all the lab work for the first time as they are a High School team. Being able to mentor them allowed us to work through the study step by step until it became second nature to perform all the readings.
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Link to Judd’s wiki on our collaboration: https://2017.igem.org/Team:Judd_UK/Collaborations
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Revision as of 02:25, 16 December 2017


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