Difference between revisions of "Team:NUS Singapore-A/shadow/Safety"

 
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   <img src="https://static.igem.org/mediawiki/2018/1/17/T--NUS_Singapore-A--Safety_header_C.png" class="header">
 
   <img src="https://static.igem.org/mediawiki/2018/1/17/T--NUS_Singapore-A--Safety_header_C.png" class="header">
  
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<div class="all-wrapper">
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   <h1>Introduction</h1>
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   <h1>OVERVIEW</h1>
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   <p>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.</p>
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   <div class="row">
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    <div class="column left">
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      <p>Clothes are a second skin - no other product is quite as intimate. We dream of our flavonoids being used to dye clothes which 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.</p>
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    </div>
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    <div class="column right">
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      <p>From our project’s inception, we have also taken great care to design a biomanufacturing process that dramatically minimizes hazards for the entire supply chain. 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.</p>
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    </div>
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  </div>
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   <br>
 
   <br>
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  <hr>
 
 
   <br>
 
   <br>
  
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   <h1>THE RIGHT CHOICES</h1>
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   <h1>Safe Project Design</h1>
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   <div class="row">
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   <p>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.<br><br>
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    <div class="column left">
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  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.<br><br>
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      <p>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 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: <a href="https://2018.igem.org/Team:NUS_Singapore-A/Design">Design</a>). 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.</p>
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    </p>
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    <br>
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    <p>Our team decided that producing natural dyes instead could be a suitable substitute for synthetic dyes because they are non-toxic, and synthetic biology has the potential to overcome the current challenges facing natural dye manufacturing, and finally make natural dyes a viable alternative to synthetic dyes.
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    </p>
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    </div>
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      <div class="column right">
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        <p>The flavonoid dyes produced in our experiments are all present in edible plants. Chrysanthemin is present in raspberries and peaches, callistephin is present in pomegranates and strawberries, and luteolin can be found in celery and broccoli. The starting reagent, naringenin, can be found in grapefruits. These flavonoids are not known allergens. Furthermore, they have historically been used as natural dyes. As such, they are unlikely to cause any harm to anyone when used on clothing, during production, or if accidentally released into the environment in large quantities.
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        </p>
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        <p>They say you are what you eat. Why not wear what you eat? (Except the meat dress. Do not wear a meat dress.)
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        </p>
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    <br>
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  </div>
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   <br>
 
   <br>
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  <hr>
 
 
   <br>
 
   <br>
  
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   <h1>LAB SAFETY</h1>
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   <h1>Safe Lab Work</h1>
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    <div class="row">
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  <p>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?<br><br>
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      <div class="column left">
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  Our completed, detailed iGEM safety form demonstrates our commitment to safety, and all of us are pleased to invite you to read it <a href="https://2018.igem.org/Safety/Final_Safety_Form?team_id=2819">here</a>.</p>
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        <p>And so, 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?
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        </p>
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      <br>
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      </div>
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      <div class="column right">
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        <p>Our completed, detailed iGEM safety form demonstrates our commitment to safety, and all of us are pleased to invite you to read it <a href="https://2018.igem.org/Safety/Final_Safety_Form?team_id=2819">here</a>.
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        </p>
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      </div>
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    </div>
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  <hr>
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  <br>
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  <br>
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  <br>
  
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    <h1>HUMAN PRACTICES</h1>
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  <h1>HUMAN PRACTICES</h1>
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    <div class="row">
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  <p>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.<br><br>
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      <div class="column left">
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  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.<br><br>
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        <p>As word of our project spread, designers like 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.
+
  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 <a href="https://static.igem.org/mediawiki/2018/0/04/T--NUS_Singapore-A--Safety_protocol.pdf" download="NUS Singapore-A Safety Protcol with Evaluation">here</a>. 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!<br><br>
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        </p>
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  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!</p>
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        <br>
+
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        <p>At that time, we were filling in the first draft of safety forms, the very same one we mentioned earlier. 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 project 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 synthetic biology.
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        </p>
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      </div>
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      <div class="column left">
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        <p>Hence, we wrote to the iGEM Safety Committee with a protocol we developed for ensuring that the product we extract is microbe-free:</p>
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        <p>We first centrifuge our bacterial culture at 5,000rpm to pellet the cells. The supernatant containing the flavonoid dyes is subsequently sterilised by filtration using the 0.22um filter. Next, we plate the filtered supernatant on LB (lysogeny broth) agar plates with and without antibiotic (which the genetically engineered bacteria are resistant to; e.g. kanamycin, chloramphenicol). This decontamination protocol ends by incubating these plates overnight in 37 degree Celsius.</p>
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        <p>The absence of colonies on both plates will indicate that our dyes are free from any microbes.<br>
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        We are proud to say that this protocol has been approved by the iGEM Safety Committee!</p>
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      </div>
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    </div>
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  <p>Here are the agar plates of our decontamination test. No colonies were observed.
 
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</p>
 
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  <figure class="figures">
 
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    <img src="https://static.igem.org/mediawiki/2018/b/bc/T--NUS_Singapore-A--Luteolin1.8.1.jpg">
 
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    <figcaption>Figure 1: <i>E.coli</i> co-transformed with Brep-FNS and Brep-F3’H plasmids are plated on LB agar with Kanamycin and Chloramphenicol. </figcaption>
 
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  </figure>
 
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  <figure class="figures">
 
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    <img src="https://static.igem.org/mediawiki/2018/d/da/T--NUS_Singapore-A--Luteolin1.8.2.jpg">
 
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    <figcaption>Figure 2: <i>E.coli</i> co-transformed with pBAD-FNS and Brep-F3’H plasmids are plated on LB agar with Kanamycin and Chloramphenicol.</figcaption>
 
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  </figure>
 
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  <figure class="figures" style="width:40%;">
 
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    <img src="https://static.igem.org/mediawiki/2018/f/ff/T--NUS_Singapore-A--Luteolin1.8.4_NoAntibiotic.jpg">
 
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    <figcaption>Figure 3: Wild-type <i>E.coli</i> BL21 star is plated on LB agar without antibiotic.</figcaption>
 
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  </figure>
 
  
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     <p>We are deeply grateful to the Safety Committee for validating our efforts towards developing a safe biomanufacturing process.</p>
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  <h1>Results</h1>
 +
  <p>Colonies were absent in both types of LB agar plates, with and without antibiotics (Figure 1 Left and Right). As we explain in our decontamination protocol, this proves that our decontaminated dye product contains neither our engineered microbes nor non-engineered microbes.</p>
 +
 
 +
  <figure class="figures">
 +
     <img src="https://static.igem.org/mediawiki/2018/4/47/T--NUS_Singapore-A--Safety-result.jpg">
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    <figcaption style="text-align: left">Figure 1 Left. Filtered supernatant from luteolin-producing E.coli culture plated on LB agar with kanamycin, 2nd August 2018. Naringenin is the substrate used in luteolin synthesis. LB+K, lysogeny broth agar plate supplemented with Kanamycin at the concentration of 50ng/ul.<br>
 +
    Figure 1 Right. Filtered supernatant from luteolin-producing E.coli culture plated on LB agar without antibiotics, 6th August 2018. Naringenin is the substrate used in luteolin synthesis.
 +
    </figcaption>
 +
  </figure>
 +
 
 +
  <h2>Safe Shipment</h2>
 +
  <p>We are pleased to report that we did not face any safety problems in sending our DNA parts to the Registry.</p><br><br>
 
    
 
    
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  <br>
 
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  <br>
 
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  <hr>
 
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  <br>
 
  
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    <h1>BIOMANUFACTURING PROCESS</h1>
 
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        <p>After approval, we were spurred on to develop a biomanufacturing process that uses as little potentially toxic, harmful, or pollutive chemicals as possible. If it could have a positive impact on the environment, even better. To do this, we explored some novel methods to increase the eco-friendliness of our biomanufacturing process.
 
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      <div class="column right">
 
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        <p>We tried to use xylose from empty fruit bunches as a feedstock, giving the waste material a new lease of life. </p>
 
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Latest revision as of 06:48, 15 October 2018

CONNECT WITH US

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

Colonies were absent in both types of LB agar plates, with and without antibiotics (Figure 1 Left and Right). As we explain in our decontamination protocol, this proves that our decontaminated dye product contains neither our engineered microbes nor non-engineered microbes.

Figure 1 Left. Filtered supernatant from luteolin-producing E.coli culture plated on LB agar with kanamycin, 2nd August 2018. Naringenin is the substrate used in luteolin synthesis. LB+K, lysogeny broth agar plate supplemented with Kanamycin at the concentration of 50ng/ul.
Figure 1 Right. Filtered supernatant from luteolin-producing E.coli culture plated on LB agar without antibiotics, 6th August 2018. Naringenin is the substrate used in luteolin synthesis.

Safe Shipment

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





HEAR OUR STORY

We’re using synthetic biology to change the way the world thinks and functions.

We’re a little disruptive, dangerous and maybe a tad too bold. But the world needs some shaking up every now and then, and we think we’re just the right people for that.

Together we’re going to engineer biology and create something awesome.

CONTACT US

nusigem@gmail.com

21 Lower Kent Ridge Rd, Singapore 119077