Alexa Dekorte and Christina Harbin

Kill switch engineering:

Our role on the team this cycle was the collaborative development of our kill switch. This included designing a plasmid, ordering the DNA, inserting the circuit into a designed plasmid backbone, transforming the plasmid into competent cells, and growing the bacteria on colony plates as well as in liquid cultures. We ran a variety of assays to assess the function and synthesis of the kill switch. This included various rounds of colony PCR followed by gel electrophoresis and sequencing the plasmid and analyzing the results. After collecting enough evidence of proper synthesis for specific colonies, we began testing the development of the bacteria under different conditions to assess the behavior of the kill switch. To test the efficacy and confirm the action of the kill switch, varying levels of HSL were used to prepare agar plates, which colonies were transplanted onto. The plates were either 10ul, 100ul, or a control plate. There were four colonies suspected to have successfully integrated the kill switch, and these were prepared to an OD of roughly .6 and then serial dilutions of 106 bacteria per uL to 103 bacteria per uL. These were then spotted onto the plates and allowed to grow. The results then showed proof of concept. The higher concentrations of cells showed some growth in a few tests but the majority of samples could not grow as the kill switch was successful in terminating cell growth when not in close proximity to high volumes of other E. Coli cells.

Matthew Hanson, Rachel Kostrzewa, and Sebastian Nolte

CFPS:

Our role on the team during this cycle was proof of concept. Over the summer, we tried to assemble the circuit of rainbow trout estrogen receptor, tetracycline repressor, and red fluorescent protein. However, though the parts were produced with the correct primers, the genes would never assemble and the cells would not express these genes. Therefore, we saw an issue occurring either with the Gibson Assembly of the genes, or there was too much burden on the cells. Therefore, we decided to test out the Cell Free Protein Synthesis to help bypass any issues with Gibson Assembly and Transformation. There were a few things that were tested. First, we wanted to see how concentration sensitive this process is by testing the cell free protein synthesis at lower concentrations. From this, we saw no red fluorescent protein being expressed meaning that this process is concentration sensitive. Lastly, we created the solutions at the correct concentrations and now are testing whether or not the process works.

George G. Amoako

Improvement of existing part:

My role on the team this cycle was to test and validate six mutations introduced into the origin of replication of pSB1C3 plasmid. Over the course of the project I have used different methods to measure the plasmid copy number of all six mutants and the wild type. I explored quantitative PCR, “special” DNA isolation and gel electrophoresis to separate the genomic and plasmid DNA. I also explored how plasmid copy number affects red fluorescent protein expression through the LAC I gene on the genome. Parts of all mutants have been created on the iGEM registry.

Connor Arens

This cycle I continued my work in the wet lab, this year working to pick up work around the different focuses of the wet lab team. I aided in preparing and testing the kill switch, by creating test cultures and colony plates to test the efficacy of said kill switch. I also worked towards contributing to our contributions to an existing part, by examining the Rainbow Trout Estrogen Receptor further. I also aided in the creation of a double inverter to prevent the RFP gene from expressing its tendency to leak and turn cells red through faint expression of the RFP gene later in cell lines.

Taylor Neeb

A Daunting History of DDT:

DDT was the first modern synthetic pesticide to be created in the 1940s. It was first employed successfully in both military and civilian populations to battle malaria, typhus, and other insect-borne human illnesses. It was also excellent in controlling insects in agriculture and animal production, institutions, houses, and gardens. Because of DDT's rapid success as a pesticide and widespread usage in the United States and other countries, many insect pest species developed resistance (DDT - A Brief History and Status). The EPA issued a cancellation order for DDT in 1972 due to its negative environmental consequences as well as its classification as a probable human carcinogen. DDT is a long-lasting organic pollutant that is easily absorbed by soils and sediments and will bioaccumulate, which can serve as both sinks and long-term sources of exposure for species (Why should DDT still be used?). We discuss the daunting history of DDT, and how the presence of DDT is still affecting ecosystems both on land and in water, chemical manufacturers, and the tobacco industry.

Mackenzie Breitner

Diversity and Inclusion initiation via a HisCO project:

This cycle our team wanted to create a partnership with Alma Colleges Diversity and Inclusion offices to show our campus community the wide range of diversity in the STEM field. To establish this partnership I worked with the Diversity and Inclusion offices as well as HisCO to create a panel called Latinx Voices in STEM. This event will showcase speakers from the Latinx community and share their experiences working in the STEM field.

Alice Hutchins

The DDT and Malaria Question:

This paper is a research and environmental ethics paper on the modern and historical issues surrounding DDT. This paper was written by Human Practices member Alice Hutchins, and helped our team learn more about the ethical issues present with the contaminant DDT, which is a central part of our project.

Human Practices Team

Stem Kit design and Book Publishing:

This cycle, work was continued towards science education, with a focus on outreach to younger audiences. This included continued work on our children's book Go Away Pollution from last year’s cycle. This year to further the engagement of younger audiences furthered our development of the STEM kit. This interactive kit contains multiple science experiments that would be a fun way to learn about synthetic biology and help younger audiences apply the work our team is doing.

MCA Writing Competition Involvement:

Human Practices produced a project implementation proposal for a grant competition through the Michigan Colleges Alliance (MCA) College Community Challenge (C3). Our goal was to engage in the ‘Design Thinking Process’ where we find a solution to a problem in our local community and tailor it to the community’s needs. This is where the idea of the STEM kit was born, as we needed a way to address the lack of prioritization of STEM classes in K-12 curriculums. Additionally, we created a card game to further foster a love for STEM in classrooms and the community.