Team:ULB-Brussels/Safety

From 2014.igem.org

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Safety
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<font color="#CCD6EA">'Some words about</font> $Safety$ <font color="#CCD6EA">in the Laboratory'</font>
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The main concerns raised by Mighty Coli depend more of the protein that we chose to produce than of Mighty Coli itself. However, Mighty Coli could compel an escaped recombinant bacterium to produce an industrial protein in the environment, when a bacterium without our system would quickly degenerate and stop producing the protein of interest. The risk seems thin, since such an overproducing bacterium would suffer from a clear competitive disadvantage in a wild environment.</p>
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The main concerns raised by Mighty Coli depend more on the protein that we chose to produce rather than on Mighty Coli itself. Mighty Coli could compel an escaped recombinant bacterium to produce an industrial protein in the environment, when a bacterium without our system would quickly degenerate and stop producing the protein of interest. The risk seems thin, since such an overproducing bacterium would suffer from a clear competitive disadvantage in a wild environment.</p>
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<p>However, in the current state of our project, the plasmids are maintained in the bacterial population through the usual system of antibiotic resistance, since the toxin and the antitoxin are to be placed on different plasmids bearing different resistance genes. The properties of the TA systems are used to boost protein production and also for plasmid stability.  Furthermore, in the final version of our project (that is, if we have the time to go this far), the toxin gene will be inserted in the genomic DNA of the bacteria, preventing it to ever be lost, and compelling the bacteria to keep the plasmid bearing the genes of the antitoxin and the protein of interest.</p>
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<p>However, in the current state of our project, plasmids are maintained in the bacterial population through the usual system of antibiotic resistance, since the toxin and the antitoxin are to be placed on different plasmids bearing different resistance genes. The properties of the TA systems are used to boost protein production but also for plasmid stability.  In the final version of our project, the toxin gene will be inserted in the genomic DNA of the bacteria to reduce loss chances. Bacteria will be compelled to keep the plasmid bearing the genes of the antitoxin and the protein of interest, without the need of infamous antibiotic resistance genes. </p>
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<p>If, for one reason or another, the overproduction of a protein did not results in a competitive disadvantage, one could also subordinate antitoxin production to an inducible promoter that would act as a built-in biocontainment device. Indeed, if we chose an artificial inducer for the expression of the antitoxin (that is, an inducer that is not found in nature), an escaped bacterium could not inhibit the activity of the toxin and woud quickly die outside of the bioreactor.</p>
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<p>If for one reason or another, the overproduction of a protein didn't result in a competitive disadvantage, antitoxin production can be put under the control of an inducible promoter, acting as a built-in biocontainment device. Indeed, if we chose an artificial inducer for the expression of the antitoxin (that is, an inducer not found in the nature), an escaped bacterium couldn't inhibit the activity of the toxin and woud quickly die outside of the bioreactor.</p>
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<p>Rules of good handling should be respected for our system.  Theses rules vary to one country to another.  
<p>Rules of good handling should be respected for our system.  Theses rules vary to one country to another.  
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According to the Belgian Biosafety Server [1], Migthy coli is in risk class 1 because our construction cannot cause any disease in animals or plants or cause disorders in the environnement. But our construction has any feedback of safety in laboratory or in industry because it’s experimental.  The risk class depends of the organism where the construction is inside but if it’s a organism of class 1 the laboratory must have different characteristic like: </p>
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According to the <a href="http://www.biosafety.be/CU/refdocs/SBB0306CU001FR.html"><b>Belgian Biosafety Server</b></a>, Migthy coli is a class 1 risk because our construction cannot cause any disease in animals or plants or cause disorders in the environnement. The risk class depends on the modified organism. To manipulate class 1 organisms, the laboratory must have different features : </p>
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<p>- A sink to wash and decontaminate the hands</p>
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- A sink to wash and decontaminate hands <br>
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<p>- A cloakroom where the lab coat are not in contact with the city coat</p>
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- A cloakroom where lab coats are not in contact with everyday clothing <br>
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<p>- The work table are easy to clean, and resistant to acidic, alkaline and disinfectant solutions</p>
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- Easy to clean benches, resistant to acidic, alkaline and disinfectant solutions <br>
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<p>- An autoclave for the sterilization of the waste</p>
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- An autoclave to sterilize waste <br>
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<p>- The lab is restricted to the personnel of the lab</p>
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- Laboratories restricted to authorized personnel <br>
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<p>- The class risk and the contact person are indicated on the entrance door of the lab</p>
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- Class risk and lab head are indicated on the entrance door of the lab <br>
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<p>- A lab coat is necessary to work</p>
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- Mandatory lab coats to work <br>
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<p>- The modified organisms are confined into a sealed system when they are not used</p>
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- Confinement of modified organisms into a sealed system when they are not used <br>
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<p>- Avoid any aerosols</p>
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- Mechanical pipetting appartus <br>
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<p>- Mechanic pipetting, not with the mouth</p>
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- Prohibiting of eating, drinking, smoking, manipulating contact lens, using cosmetics or stocking human food in the lab <br>
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<p>- Interdict of eating, drink, smoke, manipulate contact lens, use cosmetics, stock food for human consummation</p>
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- A notebook where each modified organism which is used and stocked is recorded <br>
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<p>- A notebook where each modified organism which is used and stocked is recorded</p>
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- Enforcement of lab security <br>
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<p>- The lab security is verified</p>
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- Hands are washed before leaving the lab <br>
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<p>- The hands are washed before leaving the lab</p>
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- Disinfection of benches after each manipulation and also if biological material is spilled <br>
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<p>- The work table is disinfected after each manipulation and also if biological material is reversed</p>
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- A manual of decontamination is at the disposition of the workers <br>
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<p>- A manual of the disinfectant is at the disposition of the workers</p>
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- Courses about security are applied for the workers and regular updated are organized <br>
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<p>- Courses about security are applied for the workers and a regular update is organized</p>
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- Pets are forbidden in the lab <br>
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<p>- Animals are forbidden in the lab</p>
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- Waste and contaminated materials are inactivated by an appropriate and validated method (by incineration or autoclave). </p><br>
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<p>- Waste and contaminated materials are inactivated by an appropriate and validated method (by incineration or autoclave)</p>
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<p>In conclusion, our system seems to be extremely safe and predictable. Its biggest danger consists of being used to produce a dangerous protein. This latter problem should be addressed on a case-by-case basis, by other institution than ours. Of course, our topical lab system is really safe, since it should only overproduce fluorescent reporters (GFP or RFP). However, Mighty Coli will be more useful to produce cumbersome proteins than usual fluorescent proteins.</p>
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<p>In conclusion, our system seems to be extremely safe and predictable, whose biggest danger consists of being used to produce a dangerous protein. This latter problem should be addressed on a case-by-case basis, by other institution than ours. Of course, our topical lab system is really safe, since it should only overproduce fluorescent proteins (GFP or RFP).</p>
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<p>Source : [1] http://www.biosafety.be/CU/refdocs/SBB0306CU001FR.html</p>
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Latest revision as of 19:28, 15 October 2014

$~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ \newcommand{\MyColi}{{\small Mighty\hspace{0.12cm}Coli}} \newcommand{\Stabi}{\small Stabi}$ $\newcommand{\EColi}{\small E.coli} \newcommand{\SCere}{\small S.cerevisae}\\[0cm] ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ \newcommand{\PI}{\small PI}$ $\newcommand{\Igo}{\Large\mathcal{I}} \newcommand{\Tgo}{\Large\mathcal{T}} \newcommand{\Ogo}{\Large\mathcal{O}} ~$ Example of a hierarchical menu in CSS

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- Université Libre de Bruxelles -


'Some words about $Safety$ in the Laboratory'


The main concerns raised by Mighty Coli depend more on the protein that we chose to produce rather than on Mighty Coli itself. Mighty Coli could compel an escaped recombinant bacterium to produce an industrial protein in the environment, when a bacterium without our system would quickly degenerate and stop producing the protein of interest. The risk seems thin, since such an overproducing bacterium would suffer from a clear competitive disadvantage in a wild environment.

However, in the current state of our project, plasmids are maintained in the bacterial population through the usual system of antibiotic resistance, since the toxin and the antitoxin are to be placed on different plasmids bearing different resistance genes. The properties of the TA systems are used to boost protein production but also for plasmid stability. In the final version of our project, the toxin gene will be inserted in the genomic DNA of the bacteria to reduce loss chances. Bacteria will be compelled to keep the plasmid bearing the genes of the antitoxin and the protein of interest, without the need of infamous antibiotic resistance genes.

If for one reason or another, the overproduction of a protein didn't result in a competitive disadvantage, antitoxin production can be put under the control of an inducible promoter, acting as a built-in biocontainment device. Indeed, if we chose an artificial inducer for the expression of the antitoxin (that is, an inducer not found in the nature), an escaped bacterium couldn't inhibit the activity of the toxin and woud quickly die outside of the bioreactor.

Rules of good handling should be respected for our system. Theses rules vary to one country to another. According to the Belgian Biosafety Server, Migthy coli is a class 1 risk because our construction cannot cause any disease in animals or plants or cause disorders in the environnement. The risk class depends on the modified organism. To manipulate class 1 organisms, the laboratory must have different features :

- A sink to wash and decontaminate hands
- A cloakroom where lab coats are not in contact with everyday clothing
- Easy to clean benches, resistant to acidic, alkaline and disinfectant solutions
- An autoclave to sterilize waste
- Laboratories restricted to authorized personnel
- Class risk and lab head are indicated on the entrance door of the lab
- Mandatory lab coats to work
- Confinement of modified organisms into a sealed system when they are not used
- Mechanical pipetting appartus
- Prohibiting of eating, drinking, smoking, manipulating contact lens, using cosmetics or stocking human food in the lab
- A notebook where each modified organism which is used and stocked is recorded
- Enforcement of lab security
- Hands are washed before leaving the lab
- Disinfection of benches after each manipulation and also if biological material is spilled
- A manual of decontamination is at the disposition of the workers
- Courses about security are applied for the workers and regular updated are organized
- Pets are forbidden in the lab
- Waste and contaminated materials are inactivated by an appropriate and validated method (by incineration or autoclave).


In conclusion, our system seems to be extremely safe and predictable. Its biggest danger consists of being used to produce a dangerous protein. This latter problem should be addressed on a case-by-case basis, by other institution than ours. Of course, our topical lab system is really safe, since it should only overproduce fluorescent reporters (GFP or RFP). However, Mighty Coli will be more useful to produce cumbersome proteins than usual fluorescent proteins.