Team:TU-Delft/Safety

From 2013.igem.org

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The MRSA (Methicillin-Resistant Staphylococcus aureus) problem is our inspiration for this project. This project will focus on killing Staphylococcus species with the use of antimicrobial peptides expressed in <i>E. coli</i>.
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The MRSA (Methicillin-Resistant <i>Staphylococcus aureus</i>) problem is our inspiration for this project. This project will focus on killing the Staphylococcus species by antimicrobial peptides synthesized in <i>E. coli.</i>  
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As working with MRSA requires a BSL-2 permit, and the lab in which we work has a BSL-1 permit, we did an intense literature study to find closely related organisms of same genera other than <i>S. aureus</i>. We chose <i>S. delphini</i> which has close similarities to <i>S. aureus</i>. The antimicrobial peptides where chosen in such a way that they have minimum toxicity to humans and a low minimum inhibitory concentration (MIC) for <i>S. aureus</i>.  
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As we work in the lab which has only BSL-1 permit we did intense literature study to find closely related organisms of same genera other than <i>S. aureus </i> (DSMZ 6148) that can be used for our project. We used <i>S. delphini</i> (DSMZ 20771) which had some close similarities with <i>S. aureus</i> (DSMZ 6148) to test our peptides and have a prediction of their possible effect on the MRSA . The antimicrobial peptides where chosen in such a way that they have high minimum inhibitory concentration (MIC) for humans and low for <i>S. aureus</i>.  
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The effect of antimicrobial peptides (both de-novo and natural) on humans were evaluated by testing their effects on COS 1 Kidney cells from <i>Cercopithecus aethiops</i> (African green monkey). These cells were used as they were easily accessible. The effects can be very well correlated to the effect of peptides on humans because of their shared simian nature.
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In order prevent release of live GMO's to the environment and for a safe administration of patients , we added a kill switch that is controlled by a timer. This will allow the self destruction of the cells after the desired peptides are produced. But, there are problems of release of lippopolysaccharides (LPS) after lysis, this can be overcome by using a gram positive bacterial chassis <a href="https://2013.igem.org/Team:TU-Delft/Safety#reference">[1]</a> which does not have a problem LPS.
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In order to cater to guidelines of releasing of  GMO to the environment, we added a kill switch that is controlled by a timer. This will allow the self destruction of the cells after the desired effects are attained
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<h4 align="center">Safety forms were approved on October 3, 2013 by Evan Appleton, iGEM Safety Committee.</h4>
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<a href="https://static.igem.org/mediawiki/2013/5/56/IGEM_2013_Basic_Safety_Form_signed.pdf" target="_blank">
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Safety Form 1
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<a href="https://static.igem.org/mediawiki/2013/6/63/IGEM_Biosafety_Form_Part_2_signed.pdf" target="_blank">
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Safety Form 2
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<br>
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<br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br><br>
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<a name="references"></a>
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<h2 align="center">References</h2>
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<li>Volzing, K., Borrero, J., Sadowsky, M. and Kaznessis, Y. 2013. Antimicrobial Peptides Targeting Gram-negative Pathogens, Produced and Delivered by Lactic Acid Bacteria. ACS synthetic biology.
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</li>
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Latest revision as of 13:34, 4 October 2013


Safety

The MRSA (Methicillin-Resistant Staphylococcus aureus) problem is our inspiration for this project. This project will focus on killing the Staphylococcus species by antimicrobial peptides synthesized in E. coli.

As we work in the lab which has only BSL-1 permit we did intense literature study to find closely related organisms of same genera other than S. aureus (DSMZ 6148) that can be used for our project. We used S. delphini (DSMZ 20771) which had some close similarities with S. aureus (DSMZ 6148) to test our peptides and have a prediction of their possible effect on the MRSA . The antimicrobial peptides where chosen in such a way that they have high minimum inhibitory concentration (MIC) for humans and low for S. aureus.

In order prevent release of live GMO's to the environment and for a safe administration of patients , we added a kill switch that is controlled by a timer. This will allow the self destruction of the cells after the desired peptides are produced. But, there are problems of release of lippopolysaccharides (LPS) after lysis, this can be overcome by using a gram positive bacterial chassis [1] which does not have a problem LPS.


Safety forms were approved on October 3, 2013 by Evan Appleton, iGEM Safety Committee.

Safety Form 1
Safety Form 2

References

  1. Volzing, K., Borrero, J., Sadowsky, M. and Kaznessis, Y. 2013. Antimicrobial Peptides Targeting Gram-negative Pathogens, Produced and Delivered by Lactic Acid Bacteria. ACS synthetic biology.
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