About This Project
PCB is an organic chlorine compound that bioaccumulates and persists in the environment. Production has been banned in the US, but ecosystems are still facing repercussions from decades ago. Team iTesla-SoundBio aims to combat this by transferring genes responsible for producing PCB-dechlorinating enzymes from the anaerobic Dehalococcoides mccartyi bacterium, in which they’re naturally found, into an aerobic, easier-to-work-with bacterium, such as E. coli.
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What is the context of this research?
Polychlorinated biphenyls (PCBs) are a class of man-made chemicals that contaminate many lakes and waterways. PCBs were produced for industrial and commercial applications. Although their manufacture has been banned or severely restricted, they still remain in the environment today, mainly from the disposal of old electrical equipment. Even though they persist because they are highly nonreactive and resistant to acids, bases and heat, it has been known for several decades that PCBs slowly break down in the environment. The pathway by which they are broken down was only recently discovered. A bacterium called Dehalococcoides mccartyi breaks down PCBs with a variety of enzymes, the genes for 3 of which (pcbA1, pcbA4, and pcbA5) were sequenced in 2014 by Wang.
What is the significance of this project?
PCBs are probable human carcinogens, have acute toxic effects, and cause immune system and thyroid effects. They are also known to disrupt hormone functions and have developmental effects - woman exposed to PCBs can give birth to children with significant neurological and motor control problems. Compared to other waterways, the Puget Sound has a considerably high level of PCBs, which bioaccumulate in organisms. In particular, the biomagnification of PCBs up the food chain has been severely detrimental to orcas. The bacterium Dehalococcoides mccartyi can break them down, but it is anaerobic and only obtain energy through organohalide respiration, and thus difficult to work with.
What are the goals of the project?
We plan on transferring pcbA1, pcbA4, and pcbA5 - the genes responsible for producing the PCB-dechlorinating enzymes - into E. coli or other microorganisms, as this may be very beneficial for potential PCB cleanup operations. The end goal is the development of a process using the produced enzymes to facilitate PCB cleanup or the production of technology containing the genetic pathway that could be used to clean up PCB-contaminated environments. During this, we hope to reach out to local students and the community to teach them about the importance of genetic engineering, synthetic biology, and their applications in our daily lives, as well as how they can get involved and start on their way to becoming a genetic engineer.
Here's what we'll use the funds for:
Consumable Equipment: disposable bacterial culture tubes, disposable gel extraction razors
DNA Preparation Kits: miniprep kits, biobrick kit, PCR purification kit, gel extraction kits
Gel Running Supplies: DNA ladder 100bp, DNA stain for gels
Growth Media and Supplies: LB powdered media, LB agar, chloramphenicol
Reagents and Enzymes: TAE buffer mix, T4 DNA ligase
We already have filtered tips, centrifuge tubes, PCR tubes, petri plates, DNA loading dye, DNA ladder 1Kb, ampicillin sodium, agarose, Taq DNA polymerase, high fidelity DNA polymerase, dNTP solution mix, pre-made component cells, bleach, cleaning ethanol, and glycerol, as well as custom DNA oligos.
Our sponsors are the SoundBio Lab, Fred Hutchinson Cancer Research Center's Science Education Partnership, the Baker Lab at University of Washington, the Scharenberg Lab at Seattle Children's Research Institute, New England BioLabs Inc., Integrated DNA Technologies, and SnapGene.
April, May, and June are dedicated to laboratory training and preliminary research, as well as fundraising for materials and recruiting additional mentors. We received the iGEM DNA Distribution Kit in May, will buy materials in late May and early June, and conduct experiments from June to October. Over the summer, we will also raise money for merit scholarships for travel to the Giant Jamboree in November, conduct educational outreach, and attend meetups with other teams.
Jun 01, 2017
Finish lab training
Jun 15, 2017
Jun 30, 2017
Jul 09, 2017
Sep 01, 2017
Title and abstract, track selection
Meet the Team
The schools our team members come from include Nikola Tesla STEM High School, International School, Newport High School, Skyline High School, Redmond High School, and Ballard High School. The grade level distribution is as follows: 4 freshmen, 7 sophomores, 15 juniors, and 4 seniors. We have 16 female and 14 male students. Everyone on our team has a different level of laboratory experience and we're working with that by conducting lab training in the spring.
Team iTesla-SoundBio is a team of 30 high school students representing all grades of 6 different schools across western Washington. The students are enthusiastic about synthetic biology and genetic engineering, so they have chosen to pursue laboratory research through joining the team in preparation for the International Genetically Engineered Machines (iGEM) conference in November.
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