Team:ULB-Brussels/Safety

From 2014.igem.org

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$\Tgo$he 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, but it is not excluded that the plasmid containing the $\MyColi$ system maintains itself anyway through $\small Horizontal$ $\small Gene$ $\small Transfer$ (HGT), at the expend of the wild bacteria it infects, benefiting of the proprieties of the TA system.</p>
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$\Tgo$he 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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<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 only used to boost protein production, not plasmid stability. The system will thus decay quickly if Mighty Coli bacteria wander in a wild environment, without any antibiotic. 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 . It will also prevent any reasonable chance of HGT of the Mighty Coli System.</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>
<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>
<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>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, 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:
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- A sink to wash and decontaminate the hands
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- A cloakroom where the lab coat are not in contact with the city coat
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- The work table are easy to clean, and resistant to acidic, alkaline and disinfectant solutions
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- An autoclave for the sterilization of the waste
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- The lab is restricted to the personnel of the lab
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- The class risk and the contact person are indicated on the entrance door of the lab
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-  A lab coat is necessary to work
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- The modified organisms are confined into a sealed system when they are not used
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- Avoid any aerosols
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- Mechanic pipetting, not with the mouth
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- Interdict of eating, drink, smoke, manipulate contact lens, use cosmetics, stock food for human consummation
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- A notebook where each modified organism which is used and stocked is recorded
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- The lab security is verified
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- The hands are washed before leaving the lab
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- The work table is disinfected after each manipulation and also if biological material is reversed
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- A manual of the disinfectant is at the disposition of the workers
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- Courses about security are applied for the workers and a regular update is organized
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- Animals are forbidden in the lab
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- 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, 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>
<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 : http://www.biosafety.be/CU/refdocs/SBB0306CU001FR.html</p>
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Revision as of 22:36, 10 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 -


Safety


$\Tgo$he 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.

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.

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.

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 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: - A sink to wash and decontaminate the hands - A cloakroom where the lab coat are not in contact with the city coat - The work table are easy to clean, and resistant to acidic, alkaline and disinfectant solutions - An autoclave for the sterilization of the waste - The lab is restricted to the personnel of the lab - The class risk and the contact person are indicated on the entrance door of the lab - A lab coat is necessary to work - The modified organisms are confined into a sealed system when they are not used - Avoid any aerosols - Mechanic pipetting, not with the mouth - Interdict of eating, drink, smoke, manipulate contact lens, use cosmetics, stock food for human consummation - A notebook where each modified organism which is used and stocked is recorded - The lab security is verified - The hands are washed before leaving the lab - The work table is disinfected after each manipulation and also if biological material is reversed - A manual of the disinfectant is at the disposition of the workers - Courses about security are applied for the workers and a regular update is organized - Animals 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, 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).

Source : http://www.biosafety.be/CU/refdocs/SBB0306CU001FR.html


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