Team:Evry/FrenchFrog

From 2012.igem.org

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Embryos were not fed during all the week of experiment, they grew up with their own vitellus.<br/>
Embryos were not fed during all the week of experiment, they grew up with their own vitellus.<br/>
Pictures were taken with the Zeiss stereomicroscope: SteREO Lumar V12 with the camera AxioCamMR3.<br/>
Pictures were taken with the Zeiss stereomicroscope: SteREO Lumar V12 with the camera AxioCamMR3.<br/>
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The <a href="http://2012.igem.org/Team:Evry/InjectionTuto">injection tutorial</a> explains very simply with diagram how we did injection and how take care about your embryos and tadpole. The experiment carries on 5 days, from the  unfertilized egg to a swimming tadpole at stage 48-50. The GFP (or other fluorescent protein) is expressed  few hours after the fertilization to the end of the week (see below).<br/>
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The <a href="http://2012.igem.org/Team:Evry/InjectionTuto">injection tutorial</a> explains very simply with diagram how we did injection and how take care about your embryos and tadpole. The experiment carries on 5 days, from the  unfertilized egg to a swimming tadpole at <a href="http://2012.igem.org/Team:Evry/Stages”>stage 48-50</a>. The GFP (or other fluorescent protein) is expressed  few hours after the fertilization to the end of the week (see below).<br/>
The iGEM-Evry tem say a great thanks to Dr. Nicolas Pollet, Dr. Aurore Thelie and Lena Vouillot (PhD student) who teach us how to inject embryos, take care of tadpoles and how to use their microscope. They are from <a href="http://www.issb.genopole.fr/">Institute of Systems & Synthetic Biology</a> of Evry in the <a href="http://indigene.issb.genopole.fr/">Metamorphosys</a> group.<br/><br/>
The iGEM-Evry tem say a great thanks to Dr. Nicolas Pollet, Dr. Aurore Thelie and Lena Vouillot (PhD student) who teach us how to inject embryos, take care of tadpoles and how to use their microscope. They are from <a href="http://www.issb.genopole.fr/">Institute of Systems & Synthetic Biology</a> of Evry in the <a href="http://indigene.issb.genopole.fr/">Metamorphosys</a> group.<br/><br/>
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<h2>pCS2+ GFP-aid</li> </h2>
<h2>pCS2+ GFP-aid</li> </h2>
<h3>24h after injection</h3><br/>
<h3>24h after injection</h3><br/>
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Embryos are around stage 20, neural fold is visible and the size of these "neurulas" is near 1 mm <br/>
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Embryos are around <a href="http://2012.igem.org/Team:Evry/Stages”>stage 20</a>, neural fold is visible and the size of these "neurulas" is near 1 mm <br/>
<img src="/wiki/images/c/c7/409GFP-aid%2Bcontrol.jpg" alt="perdu" width="880px"/><br/>
<img src="/wiki/images/c/c7/409GFP-aid%2Bcontrol.jpg" alt="perdu" width="880px"/><br/>
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<h3>48h after injection</h3><br/>
<h3>48h after injection</h3><br/>
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Embryos are at stage 34-38 and move by intermittence, the size of tadpole is near 2.5 mm<br/><br/>
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Embryos are at <a href="http://2012.igem.org/Team:Evry/Stages”>stage 34-38</a> and move by intermittence, the size of tadpole is near 2.5 mm<br/><br/>
<img src="/wiki/images/f/f8/509_gfpaid_1et2.JPG" alt="perdu" width="880px" /><br/>
<img src="/wiki/images/f/f8/509_gfpaid_1et2.JPG" alt="perdu" width="880px" /><br/>
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<h3>Three days after injection</h3><br/>
<h3>Three days after injection</h3><br/>
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Embryos are at stage 41-42 and swim, the size of tadpole is near 4 mm.<br/><br/>
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Embryos are at <a href="http://2012.igem.org/Team:Evry/Stages”>stage 41-42</a> and swim, the size of tadpole is near 4 mm.<br/><br/>
From this tadpole stage an anaesthetic is required to take pictures of tadpoles, otherwise the light teases tadpoles, and it is difficult to take a good picture.
From this tadpole stage an anaesthetic is required to take pictures of tadpoles, otherwise the light teases tadpoles, and it is difficult to take a good picture.
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<h3>Four days after injection</h3><br/>
<h3>Four days after injection</h3><br/>
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Embryos are at stage 45-46 and swim, the size of tadpole is near 5 mm.<br/><br/>
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Embryos are at <a href="http://2012.igem.org/Team:Evry/Stages”>stage 45-46</a> and swim, the size of tadpole is near 5 mm.<br/><br/>

Revision as of 16:52, 25 September 2012

The Evry iGEM team is proud to introduce you to our new project: The French Froggies!

The French froggies project

Establishment of a new chassis



So far, synthetic biology has mostly focused on bacteria, since they are simple to engineer. iGEM teams and laboratories have worked on unicellular organisms in order to understand the underlying biology and have developed an impressive database of molecular parts. Some work has also been done on engineering mammalian cells and a few iGEM teams have followed this trend. Synthetic biologists are now imagining the rational design of multicellular organisms with numerous applications ranging from gene therapy or drug production to environmental monitoring. This year, our team would like to be part of that challenge.

The arrival of Xenopus as a chassis in synthetic biology requires the creation of new standards and protocols that the community will be able to build on. We provided the registry with such tools that allow rapid construction and characterization of devices in vivo, and include debugging tools. We think they will be very useful for later iGEM teams and synthetic biologists who wish to work with Xenopus for building multicellular systems.


You want to make the move from bacteria to multicellular synthetic biology ? Make sure you check out our Introduction to Xenopus page, and our Frogs for dummies page to make sure you are aware of all the differences between genetic engineering in eukaryotes

This year, the Evry iGEM team is going to be the one of the first iGEM team to work on a vertebrate. Our work is focused both on developing a system for intercellular and inter-tissue communication, and creating the tools for the iGEM community to easily express genes in specific tissues. We believe the tadpole is a chassis of choice for iGEM on multi cellular organisms, as experiments can be conducted in one week using microinjection methods. We hope to demonstrate the feasibility of engineering Xenopus in one summer for an iGEM project, and to create a great tool for multicellular synthetic biology: A synthetic, orthogonal hormonal system.


The simple molecular strategy to build eukaryotic plasmid ready to use:




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Characterization of one injected plasmid into Xenopus tropicalis eggs for a week