Poster: FDR-HB_Peru

Fishing for Cadmium

Members

Daniela Gioia¹, Miski Nopo², Isabela González¹, Kia Cuba¹, Andrew Fenton¹, Gabriel de Romaña¹, Sofia Villafuerte¹, Ga-Hyun Kim¹, Thiago Aguirre², Jorge Alonso Diaz¹, Ben Falls¹, Marry Xuan¹, Santiago Ochoa², Rafaella Velasquez¹, Alejandra Domínguez², Alejandra Reategui², Valeria Miranda², Romina Barrantes², Simon Cubas-Koyama¹, Antonia Portocarrero², Isabella Yoshiyama¹, Celine Hoyle¹, Kade Webber¹, and Miguel Garro²

Mentors

Dr. Nina Markham¹, Dr. Claudia Muñoz², and Mr. David Kostial¹

¹ = Colegio Franklin Delano Roosevelt

² = Hiram Bingham the British International School of Lima

Abstract

Our project seeks to tackle the issue of cadmium contamination in fishmeal. We chose this issue because it affects the health of humans, ecosystems, and our local economy. To tackle this issue we have designed a plasmid capable of detecting the concentration of cadmium in blended fish. We have plans to implement a cell-free system to make our solution more accessible to fishermen. We are also working towards making an affordable heated shaking incubator to make the culturing of bacteria easier for other iGEM teams and people working in synthetic biology. Lastly, we have taken strides to make science education more accessible for young adults in Peru, by creating videos, evaluating articles for a young science journal, and subtitling our previous TEDx talk.

Inspiration

About three years ago, our iGEM team visited the fishing company TASA, at one of their fishmeal factories. We found that the fishing industry is greatly affected by heavy metal, specifically cadmium, contamination caused by illegal gold mining in the Amazon. We continue to consistently work with TASA and our collaboration continues to inspire our work.

Problem

Cadmium is a prevalent contaminant in Peru, affecting peruvian soil, and water. Through the food chain humans ingest cadmium which can lead to lung, kidney, and neurological damage. Cadmium in exported products, such as fishmeal and cacao, decreasing their competitive edge therefore negatively impacting our local economy.

Idea

Last year we focused on making a bioassay that used live bacteria to detect different concentrations of cadmium. We changed our focus towards a cell free system because it would be easier for fishermen to use and would decrease our detection time.

Integrated Human Practices

We have been working with TASA, a large fishmeal company, for two years. Our goal has been to develop a Cadmium (Cd) detection system for them.

In discussions with TASA, we confirmed that fishermen probably won't feel comfortable working with bacteria on the boat and presented our idea for a cell-free system.TASA agreed that we should pursue the cell-free option. Thus, we are currently designing a dipstick for the fisherman to use to detect Cd on the boat. A benefit of this option is that it could decrease the detection time of Cd from 8-24 hours down to 1-2 hours.

Meeting with TASA

Constructs

Construct 1

(BBa_K3466000)

The MerR protein, constitutively expressed, represses the PcadA promoter. When cadmium is present, the MerR protein is released from the PcadA promoter site, allowing RFP to be expressed, signifying the presence of Cadmium.

Construct 2 and Construct 3

(BBa_K3466001)

(BBa_K3466002)

Spacers were added before the PcadA promoter in Construct 2 and after the PcadA promoter in Construct 3 to test whether or not they serve to enhance our RFP expression rate.

Construct 4, Construct 5, and Construct 6

(BBa_K3466003)

(BBa_K3466004)

(BBa_K3466005)

(BBa_K3466006)

These plasmids are versions of constructs 1, 2, and 3 with the addition of BamHI and BgIII restriction sites surrounding the RBS to allow for the insertion of additional RFP segments if necessary for detection.

Construct 7

(BBa_K3466007)

Construct 7 is an alternate version of construct 1 with the substitution of a T7 promoter. This should increase efficiency in our cell-free system.

Parts Used

T7 promoter - BBa_K3033000
Constitutive promoter - BBa_J23100
RBS+MerR - BBa_K1724002
PcadA promoter - BBa_K1724000
RBS - BBa_J34801
mRFP - BBa_I13521
TT - BBa_B0015

Experimental Design

Our goal is to create a paper-strip style dipstick test that will produce a visible color change when certain levels of Cadmium is present. This flowchart represents our experimental design in order to achieve this goal.

Hardware

Shaking Incubator - Open-Source Manual

When realizing that Africa and South America accounted for less than 4% of the teams, we couldn't help but question the financial aspect of the situation. To bridge this gap, we have worked on turning our last year's hardware device (bioassay) into an open-source instruction manual for the construction of the shaking incubator to help other institutions have an easy and cheap way of growing bacteria. What makes this even better is that it can help people all over the world practice synthetic biology as long as they have $40 (reduced from common $900).

Cell-Free Test

We have started to create initial freeze-drying protocols to be carried out when possible to turn our liquid cell-free system into a dipstick fast test. This would make our solution easier to use (no onsite pipetting, less maintenance, etc.).

Education & Outreach

TEDxYouth@Tukuy 2020 Workshop

On August 22, TEDxYouth@Tukuy hosted an event for 700 students from all over Peru, where two of our team members led a webinar about synthetic biology and problem solving.

TEDx Talk: “La ciencia en manos de escolares”

This year, our iGEM team’s TEDxTukuy talk was uploaded on Youtube and to the official TED website. In September, we created subtitles for this TED talk both in English and Spanish and submitted them to the TED organization for them to be uploaded to the TEDx talk video. These subtitles will make the video more accessible to a worldwide audience.

Educational videos produced in collaboration with UPCH_Peru

In collaboration with the UPCH iGEM team we created a series of scientific educational videos for Peruvian students, who had limited access to science education.

The series was made up of 4 parts in total:

Introduction to the scientific method
Introduction to synthetic biology
FDR-HB_Peru team project
UPCH_Peru team project

Workshop organized in collaboration with UPCH_Peru

We organized a hands-on virtual event for youth from three main regions of Peru held on October 24th alongside the UPCH_Peru iGEM team. This event had the goal of informing students about synthetic biology and our team projects, but most importantly developing their curiosity and interest in science.

Journal Club Peru Presentation

We gave an online presentation on Journal Club Peru’s Facebook platform with over 10,000 followers. Three of our iGEM members explained the basics of synthetic biology and described our project in a fun and engaging way for other Peruvian students.

Frontiers for Young Minds

Frontiers for Young Minds is a science journal for young students, reviewed by middle and high school students before they get published. We help provide feedback to scientists on how to modify their articles to make them more appealing and understandable to young audiences.