Team:GeorgiaTech/Collaborations

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Collaborations

Lambert High School

On July 14th, our iGEM team hosted a collaboration on Georgia Tech’s campus with the Lambert High School iGEM team. In the morning, the Lambert High School students presented their project to us. Afterwards, we had a discussion about their project and gave them a lot of suggestions about how to proceed with the final few weeks before their fall semester began again.

To summarize their project, they wanted to develop a product that could be used to detect early stages of bacterial infections in farmers’ crops by employing a fluorescent green fluorescent protein (GFP) indicator tag. Their project consisted of three components. First, they wanted to use a CRISPR/Cas 9 system to detect the infectious bacteria. The system would involve a strand of guide RNA (gRNA), complementary to a gene unique to the bacteria, so it would be able to identify the presence of the bacteria and signal the CRISPR to cut the DNA of the bacteria. During the subsequent DNA repair stage, a GFP indicator gene would be incorporated through a “knocked-in”. The second part of their project addressed the characterization of the GFP-labeled crops. They wanted to visualize of the fluorescent indicators from the GFP knock-in using a modified version of Foldscope microscopy, a more affordable imaging technique that would be accessible to high school students. The third part of their project involved expressing chitosan deacetylase (CDA) in E. coli. Since CDA is toxic to E. coli, they designed a plasmid with promoters, RBSs, GFP, Pelb, and CDA genes on ApE software and asked for our suggestions before they order the gene from IDT.

After listening to their project proposal, We also suggested that they focus the next 8 weeks of their project on only the CDA system, since the CRISPR/Cas 9 system is more difficult to work with. As shown below, we proposed three different constructs for the CDA vector that would express both the CDA and Pelb genes.

Our suggested constructs are as follows:

1. An bicistronic system with ORFs going in the same direction
2. A fusion protein headed for the periplasm
3. A bidirectional system with 2 ORFs that would go in opposite directions and control 2 different pieces of mRNA

We also suggested that they make the best use of their time by approaching the project from all three fronts by dividing their team into three groups, each dedicated to working with one of the three constructs.

After discussing Lambert High School’s project, we presented our project to them and taught them about the central concept, including click chemistry, phage display, and mutagenesis. We also gave them a tour of our lab and showed them all the instruments that we use.

We spent the whole day with the Lambert High school iGEM team, and our mentors had time to interact and chat about their research projects. At the end, we set up a date for a future meeting where we will both give our finalized presentations as practice for the Giant Jamboree.

We kept in touch with Lambert during the summer, and by September, the Lambert High School iGEM team had successfully synthesized the three constructs and used them to test their CDA vector. More information about their project can be found at their team Wiki page.