Introduction

Clothes are like a second skin - no other product is quite as intimate. We dream of producing flavonoids to dye clothes that are beautiful and meaningful; to create an everyday magic within anyone’s reach. It is thus our top priority to ensure that our dyes, which will be in constant contact with one’s skin, is safe for everyone to use. In fact, we ought to ensure that for any product birthed from our biomanufacturing platform. At our project’s inception, we also took great care to choose safe and beneficial compounds to produce. We present to you our answer to the toxic practices of the fashion industry today, and the story of how we built our culture of safety.

Safe Project Design

Beauty comes at an ugly cost. The fashion industry is the second largest polluting industry in the world, second only to fossil fuel energy production. According to Mr. Vinod Agnihotri of LANXess, the most toxic part of the fashion industry is the manufacturing of synthetic dyes. Not only does it take a toll on factory workers’ health, but also the effluent is often discharged into nearby water bodies, harming the local flora and fauna, as well as severely decreasing the quality of life of everyone living nearby. It is difficult to get rid of synthetic dyes once they are released into the environment because they have been designed to be extremely persistent. It feels great when our clothes still look brand-new after a long time, but it’s bad news for the environment.
Our team decided that producing natural dyes instead could be a suitable substitute for synthetic dyes because they are non-toxic, and synthetic biology, especially optogenetics, has the potential to overcome the current challenges facing natural dye manufacturing, and finally make natural dyes a viable alternative to synthetic dyes.
Our starting reagent, naringenin, can be found in grapefruits. Our product, luteolin, is found in celery, thyme, green peppers, and chamomile tea. It is not a known allergen - in fact, it has an anti-allergic action ! Furthermore, it has historically been used as a natural dye (See: Design). As such, our compounds are unlikely to cause any harm to anyone when used on clothing, during production, or if accidentally released into the environment in large quantities.

Safe Lab Work

The time came for us to start developing a synthetic biology solution to this problem. We knew that it was of fundamental importance that we adhere to lab best practices because if we can’t follow safe microbiological lab procedures, and prove that it is possible to produce our dyes safely, how can we expect anyone to be convinced by our solution?
Our completed, detailed iGEM safety form demonstrates our commitment to safety, and all of us are pleased to invite you to read it here.

HUMAN PRACTICES

As word of our project spread, designers like Miss Leong Minyi expressed interest in using our dyes. We were reminded that we needed to consider the future of our project and how it would impact society.
At that time, we were filling in the first draft of safety forms, the very same one in the previous section. However, our excitement to share our project with the world was dampened when we realized that the iGEM safety rules stated that we were not allowed to release any products derived from our work in the lab. Nevertheless, we firmly believed that our luteolin deserved to be released, so that we could get useful and important feedback from our targeted end users, designers, as well as show the world the revolutionary potential of optogenetics in biomanufacturing.
Hence, we wrote to the iGEM Safety Committee with a safety workflow we developed for ensuring that the product we extract is microbe-free. You may download it as a PDF here. This document also includes a guide on how to obtain permission from the iGEM Safety Committee to safely bring biomanufactured products out of the lab. This was done in the hope that future teams will find our story helpful and inspiring, and be more aware of biosafety and security issues and best practices. It is our dream that more biomanufactured products can be appreciated and celebrated at future Jamborees!
We are proud to say that our workflow was approved by the iGEM Safety Committee! See our results below, and visit us at the Jamboree to see our dyes and find out more!

Results

Figure 1. Filtered supernatant from luteolin-producing E.coli culture plated on LB agar with kanamycin and chloramphenicol, 30th September 2018.

To further prove the effectiveness of our decontamination protocol, we plated supernatant from the wild-type E.coli BL21 Star (DE3) culture on LB agar without antibiotics, and incubated it overnight. Again, colonies were absent (Figure 2), showing that our decontamination protocol works on non-engineered microbes.

Safe Shipment

We are pleased to report that we did not face any safety problems in sending our DNA parts to the Registry.