Team:UNebraska-Lincoln/Integrated Practices 1
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How Human Practices were Integrated into our Project:
Human practices directly contributed to our awareness of biosafety. We discussed our project and safety concerns with scientists from Monsanto, Li-Cor, and Nature Tech. The feedback we recieved from them was valuable. Since our engineered microorganism is inended to be introduced into a marine environment, scientists from Nature Tech suggested bacterial conjugation should be a primary concern. Incorporating the genes onto the chromosomal DNA could mitigate the concerns with bacterial conjugation. Our project is only a proof of concept, actually adding our gene of interest onto the chromosomal DNA is out of the scope of our project, but it was still a consideration when designing and developing safety cases.
This summer, our team attended the Heartland iGEM Regional Meetup. Here, we participated in a Building with Biology public forum on engineering Mosquitos to fight malaria and helped host a BioBuilders workshop to help educate students and the community. It became very clear to us that biosafety and bioethics were some of the community's main concerns.
This regional meetup and the previous discussions with scientists from biotech companies affirmed the importance of safety in synthetic biology. Our team decided to make safety one of our project’s centerpieces. The safety case we developed is a unique exemple of how we integrated safety concerns we became aware of from Human Practices. We hosted our own Building with Biology Public event at a local museum where we educated public on synthetic biology as a whole and emphasized safety measures that are currently used in synthetic biology. Developing a relationship with the public and building safety cases played a large role in our project.
How did safety discussions influence our design?
Through safety discussion, our team realized the importance of having a kill switch. Our engineered microorganism is intended to reduce nitrate ions in marine environments. The introduction of nitrate reducing E. coli into waterways could have unintended environmental impacts. We decided to develop a kill switch. We identified environmental concerns with many kill-switches and designed a kill switch with a light environmental footprint.
Being mindful of safety, we decided to use a strain of E. coli that was auxotrophic for serine as our chassis. A plasmid allowing for the synthesis of serine that is regulated by a nitrate sensitive promoter was developed. We decided to use a chassis auxotrophic for serine for several reasons:
- Most every organism requires the amino acid serine for survival
- Natural marine environments lack the concentration of serine required for survival of our chassis
- Many other kill switches induce death through synthesis of toxins
- If other bacteria uptake our kill switch through bacterial conjugation, they will synthesize more serine in the presence of nitrates, consequences will be minimal; serine is already synthesized in bacteria
More details of our kill switch design can be found here.
