Team:EPF Lausanne/Project

From 2010.igem.org

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= Overview =
= Overview =
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This page provides an overview of our project and the [[Team:EPF_Lausanne/Project#Results|results]] we achieved.
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==Project idea==
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[[Image:Mousquito and asaia 3.png|left|250px|caption]]
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The aim of our project is to defeat [https://2010.igem.org/wiki/index.php?title=Team:EPF_Lausanne/Project_malaria Malaria]. To do so, we genetically modified a bacterium that is naturally present in ''Anopheles'' mosquitoes. Our engineered bacterium is designed to prevent malaria from infecting the mosquito. This in turn will prevent the transmission of the parasite to humans.
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Our projects aims to help stopping the propagation of malaria. To do so, we chose to act on the mosquito. Indeed, if we prevent the mosquito from being infected by malaria, it won't be able to transmit the parasite to healthy humans.
 
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Our idea is to engineer asaia, a bacteria that is naturally present in the mosquito's intestinal tract. This bacteria would express an immunotoxin to kill the plasmodium and thus prevent the mosquito infection.
 
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= Movie =
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Our idea is to engineer [https://2010.igem.org/Team:EPF_Lausanne/Project_asaia ''Asaia''], a bacterium that is naturally present in the mosquito's intestinal tract.
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Coming soon!
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[[Image:Asaia_project_overview.png|left|300px]]
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[[Image:gut.png|300px]]
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=Plan=
 
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After choosing our bacteria of interest, here Asaia, we decided on two main set of experiments.
 
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*Cloning
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This bacterium would express an [https://2010.igem.org/Team:EPF_Lausanne/Project_immuno immunotoxin] or specific [https://2010.igem.org/Team:EPF_Lausanne/Project_immuno proteins] that either kill ''P. falciparum'' or prevent its entry into the epithelium of ''Anopheles''.
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The first of course, was the cloning part. We performed all the cloning in E.coli, and did experiments on Asaia transformation, growth and resistance to antibiotics in parallel.
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Here is the parts we successfully cloned.
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As E.Coli origin does not work in Asaia, we added an origin that was compatible with this bacteria. ''Gwen: where did we get this origin from?'' On the opposite, Asaia's origin does work in E.Coli, which is very useful for our sets of experiments.
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*dessin des "parts"*
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[[Image:all_team.png|200px|caption]]                [[Image:hunting.png|200px|caption]]
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*Drosophila
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Our host bacterium ''Asaia'' is an organism that is not only easy to grow and to genetically manipulate but is also naturally present in the ecosystem. We aim to establish ''Asaia'' as a new chassis to give future iGEM teams the opportunity to quickly and efficiently engineer new and more potent ''Asaia'' strains. This would provide synthetic biology with a useful tool in the fight against malaria and other mosquito borne diseases.
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In parallel, we did experiments to assess if Asaia was persistant in ''Drosophila melanogaster ''. In fact, flies are much easier organism to grow and work on, in opposition to mosquitoes which need to feed on animals to reproduce.
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[[Image:Zorro.png|right|300px|caption]]
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Our experiment was set in 3 steps:
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*infection of the flies with GFP-Asaia*, and 3 other bacteria as controls
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*retrievement of the flies after 3h (control of infection), 24h, 48h
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*counting of the number of colonies for all time points and the 4 differents strains
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The goal was to observe the persistance of Asaia in the Drosophila gut to see if it can be used as a model organism to study Asaia.
 
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So... ''ASAIA'' is the pink power against malaria.....
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For an overview of our project, you can watch our movie
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[https://2010.igem.org/Team:EPF_Lausanne/Humanpractices_Movie iGEM EPFL movie]
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[[Image:Movie chair.png|center|100px|caption]]
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==Results==
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[[Image:EPFL check list results.png|right|380px|caption]]
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We established [https://2010.igem.org/Team:EPF_Lausanne/Project_asaia ''Asaia'' as a new chassis] by providing detailed [[Team:EPF_Lausanne/Project_asaia#Do you want to work with Asaia?|tech-sheets]] on how to manipulate it.
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We constructed several [https://2010.igem.org/Team:EPF_Lausanne/Project_parts biobricks] with which [https://2010.igem.org/Team:EPF_Lausanne/Project_immuno proteins] can be produced that may block [https://2010.igem.org/wiki/index.php?title=Team:EPF_Lausanne/Project_malaria malaria] infection. We also made Asaia-specific biobricks, such as the [[Team:EPF_Lausanne/Project_parts#Promoter_characterization|CFP construct to test expression]].
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These tools will allow future iGEM teams to use ''Asaia'' and manipulate it
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to fight mosquito-borne diseases.
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The [[Team:EPF_Lausanne/Project_immuno#Results|expression of the proteins]] intended to block ''P. falciparum'' was tested in ''E. coli''. The tests showed that the immunotoxin is expressed and it also appears in the supernatant, which is evidence for secretion.
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We could not show expression of the P-proteins, probably due to the AT-rich sequence.
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Furthermore we tested the [[Team:EPF_Lausanne/Project_droso#I) Experiments on Drosophila|persistence of ''Asaia'' in ''D. melanogaster'']] to see if it could be used as an alternative host to facilitate research. The results showed that ''Asaia'' is not persistent in ''D. melanogaster'' and we assume that it is because ''Asaia'' is very specific to mosquitoes.
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We also established a [[Team:EPF_Lausanne/Project_droso#II) Experiments on mosquitoes|collaboration with a Laboratory at the Pasteur Institute]] in Paris that works on mosquitoes to help us test whether our constructs indeed inhibit ''P. falciparum'' from infecting mosquitoes.
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Latest revision as of 23:18, 27 October 2010



Overview

This page provides an overview of our project and the results we achieved.

Project idea

caption

The aim of our project is to defeat Malaria. To do so, we genetically modified a bacterium that is naturally present in Anopheles mosquitoes. Our engineered bacterium is designed to prevent malaria from infecting the mosquito. This in turn will prevent the transmission of the parasite to humans.


Our idea is to engineer Asaia, a bacterium that is naturally present in the mosquito's intestinal tract.

Asaia project overview.png

Gut.png


This bacterium would express an immunotoxin or specific proteins that either kill P. falciparum or prevent its entry into the epithelium of Anopheles.

caption caption


Our host bacterium Asaia is an organism that is not only easy to grow and to genetically manipulate but is also naturally present in the ecosystem. We aim to establish Asaia as a new chassis to give future iGEM teams the opportunity to quickly and efficiently engineer new and more potent Asaia strains. This would provide synthetic biology with a useful tool in the fight against malaria and other mosquito borne diseases.

caption


So... ASAIA is the pink power against malaria.....





For an overview of our project, you can watch our movie iGEM EPFL movie

caption

Results

caption

We established Asaia as a new chassis by providing detailed tech-sheets on how to manipulate it. We constructed several biobricks with which proteins can be produced that may block malaria infection. We also made Asaia-specific biobricks, such as the CFP construct to test expression. These tools will allow future iGEM teams to use Asaia and manipulate it to fight mosquito-borne diseases.

The expression of the proteins intended to block P. falciparum was tested in E. coli. The tests showed that the immunotoxin is expressed and it also appears in the supernatant, which is evidence for secretion. We could not show expression of the P-proteins, probably due to the AT-rich sequence.

Furthermore we tested the persistence of Asaia in D. melanogaster to see if it could be used as an alternative host to facilitate research. The results showed that Asaia is not persistent in D. melanogaster and we assume that it is because Asaia is very specific to mosquitoes.

We also established a collaboration with a Laboratory at the Pasteur Institute in Paris that works on mosquitoes to help us test whether our constructs indeed inhibit P. falciparum from infecting mosquitoes.


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