Eliminating PCB pollution in the Puget Sound by genetically modifying bacteria

SoundBio Lab
Woodinville, Washington
Biology
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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.

Budget

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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

DNA Sequencing

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.

Endorsed by

This team of high schoolers is very enthusiastic, talented, and passionate about leveraging biology to solve real-world problems. Their work in this project will make meaningful steps towards solving a high impact environmental problem. The findings from this work could be applied to accelerate the rate at which this toxic chemical is removed from waterways around the globe.

Flag iconProject Timeline

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

Purchase materials

Jun 30, 2017

Project description

Jul 09, 2017

Project Launched

Sep 01, 2017

Title and abstract, track selection

Meet the Team

Team iTesla-SoundBio
Team iTesla-SoundBio

Affiliates

Team iTesla-SoundBio is collaborating with members of UW's iGEM team, SoundBio members, and other local professionals in relevant fields of study.
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Team Bio

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

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.

Lab Notes

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