Difference between revisions of "Team:Georgia State/Description"

 
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<h1>Project Inspiration and Description </h1>
 
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<h3>NEW: Bronze Medal Criterion #4</h3>
 
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<p> The inspiration behind our project was the Netflix documentary “Chasing Coral.”
 
  
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At a glance we want to transform the microalgal symbiote of coral, Symbiodinium to overcome the overarching issue of coral bleaching. We first did some research into seeing what causes corals to bleach and consequently die off. As it turns out, there isn’t just one reason but coral bleaching is a result of a multitude of different anthropogenic dependent environmental issues. But before we can even think about what gene we’d like to introduce to combat coral bleaching, we need to look to see if there have previously been any attempts to transform Symbiodinium. We found that there have been two successful transformations of symbiodinium, one in 1998 and another in 2015 however after transformation the cells were left unable to reproduce or photosynthesize. We reached out to the lab that successfully transformed symbiodinium in 2015 and asked them if we could use their plasmid construct pCB302-gfp-MBD, even though it was not optimized to be used in a dinoflagellate. We found another paper “Nuclear gene transformation in a dinoflagellate” where they assembled a dinoflagellate optimized plasmid (DinoIII) and successfully transformed it into Oxyhrris marina. We plan to use their plasmid and replace the green fluorescent protein gene with a codon optimized red fluorescent protein gene. In the lab we plan to culture Oxyhrris marina (a heterotrophic dinoflagellate), Symbiodinium microadriaticum, and Dunaliella tertiolecta (the food source for O. marina). Then we will harvest the Symbiodinium cells and attempt to repeat the transformation done in “Heterologous DNA Uptake in Cultured Symbiodinium spp. Aided by Agrobacterium tumefaciens” with equivalent or better transformation efficiency to generate a successful transformation protocol for Symbiodinium. Once we have created the DinoIII plasmid with codon optimized rfp, we will transform it into Symbiodinium as well. </p>
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<p>See how other teams have described and presented their projects: </p>
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                        <!--<h2>Our Inspiration</h2>-->
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                        <span>Chasing Coral</span>
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<li><a href="https://2016.igem.org/Team:Imperial_College/Description">2016 Imperial College</a></li>
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<li><a href="https://2016.igem.org/Team:Wageningen_UR/Description">2016 Wageningen UR</a></li>
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<li><a href="https://2014.igem.org/Team:UC_Davis/Project_Overview"> 2014 UC Davis</a></li>
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<li><a href="https://2014.igem.org/Team:SYSU-Software/Overview">2014 SYSU Software</a></li>
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                <div class="service-text"> <h2>Our Inspiration</h2>The inspiration behind our project was the Netflix documentary <i>Chasing Coral</i>. From this documentary
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we learned about the extent of the devastation to coral reefs caused by bleaching. Many of the
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scientists seen in the film expressed despair over the seeming inevitability of the destruction. While
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current efforts to protect reefs have focused on enacting marine protected areas to prevent commercial
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fishing and industrial activities from harming the reefs, these efforts cannot stop the environmental
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effects of climate change on ocean waters. In the absence of a realistic proposal to halt global warming
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by eliminating fossil fuel consumption, marine heatwaves will become more frequent and even more
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severe ( Nature Communications <strong>volume 9</strong>, Article number: 1324 (2018),
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<a href="https://www.nature.com/articles/s41467-018-03732-9">https://www.nature.com/articles/s41467-018-03732-9</a>) leading to a corresponding increase in coral
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bleaching and reef loss. About half of the Great Barrier Reef died after the heat wave of 2016/2017.<a href="https://www.newscientist.com/article/2198318-the-great-barrier-reef-is-losing-its-ability-to-recover-
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from-bleaching/">https://www.newscientist.com/article/2198318-the-great-barrier-reef-is-losing-its-ability-to-recover-
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from-bleaching/</a>.And according to the UN Special Report on Global Warming, all global reefs around
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the world are expected to completely die off if global temperatures increase by another 2 degrees
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Celsius (<a href="https://www.ipcc.ch/2018/10/08/summary-for-policymakers-of-ipcc-special-report-on-global-
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warming-of-1-5c-approved-by-governments/">https://www.ipcc.ch/2018/10/08/summary-for-policymakers-of-ipcc-special-report-on-global-
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warming-of-1-5c-approved-by-governments/).</a></div>
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  <img src="https://static.igem.org/mediawiki/2019/d/d1/T--Georgia_State--Coral5_color.png">
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<h3>Advice on writing your Project Description</h3>
 
  
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We encourage you to put up a lot of information and content on your wiki, but we also encourage you to include summaries as much as possible. If you think of the sections in your project description as the sections in a publication, you should try to be concise, accurate, and unambiguous in your achievements.
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                    <h2>The Problem</h2>
  
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                    Coral bleaching, the loss of algal symbionts necessary for the survival of cnidarian reef organisms, is a disastrous environmental issue with global consequences. No single factor has been established as the cause of this catastrophe, but there are a multitude of suspects including increased greenhouse gas emissions and rising seawater temperatures<sup>[1]</sup>.
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                        <a href="https://www.youtube.com/watch?v=u7emUF2JQx8" class="video-play-btn wow bounceInDown" data-wow-delay="200ms" style="visibility: visible; animation-delay: 200ms; animation-name: bounceInDown;"><i class="fa fa-play"></i></a>
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<h3>References</h3>
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<p>Literature
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            <div class="service-thumbnail bg-img jarallax" style="background-image: url(../wiki/images/2/2d/T--Georgia_State--31.png);"></div>
  
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Heterologous DNA Uptake in Cultured Symbiodinium spp. Aided by Agrobacterium tumefaciens
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DOI:10.1371/journal.pone.0132693
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                    <h2>The Solution</h2>
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                    Although there is no absolute certainty about the cause of coral bleaching, increasing water
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temperatures have been directly correlated with subsequent bleaching events. And coral bleaching can
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be stimulated in aquarium corals by raising the water temperatures. There are some strains of corals
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and their symbiotic algae that prove somewhat more resistant to bleaching than others. So we
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proposed to identify those characteristics that might improve the ability to withstand bleaching and use
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a synthetic biology approach to modify the algae and restore them to corals.
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<div class="row-align-center"><div class="col-6"><img src="../wiki/images/c/c4/T--Georgia_State--symbio5.png"></div></div>
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Nuclear gene transformation in a dinoflagellate
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doi: https://doi.org/10.1101/602821
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                    <h2>The Plan</h2>
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We are establishing both culturing and transformation protocols for these microalgae symbionts. We began by optimizing culturing techniques for Symbiodinium microadriaticum, Oxyrrhis marina (our model organism), and Dunaliella tertiolecta (the food source for O. marina).
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To identify optimal algal growth conditions, we tested various factors such as growth media, light intensity, and temperature. We designed a codon optimized red fluorescent protein part that was cloned into a dinoflagellate-optimized expression plasmid (DinoIII)<sup>[2]</sup> for transformation into our model organism as a proof of concept. In parallel, we are also attempting to replicate the only known successful transformation of Symbiodinium using an Agrobacterium tumefacien co-culture carrying a binary vector, pCB302-GFP-MBD<sup>[3]</sup>, and designing/executing various electroporation protocols. A genomic analysis of clade D Symbiodinium, a clade associated with higher resistance to bleaching but diminished coral growth, will identify target genes related to bleaching resistance for transformation into the growth-favorable clade C of Symbiodinium. The corals will then uptake the modified host algae, increasing their resistance to bleaching.
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<img src="https://static.igem.org/mediawiki/2019/2/20/T--Georgia_State--symbio4.png">
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<li><cite>[1]  Douglas (2003, April 4). Coral Bleaching--How and Why doi:10.1016/S0025-326X(03)00037-7/journal. </cite></li>
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<li><cite>[2]  Sprecher,B., Zhang,H. & Lin, S. (2019, April 9). Nuclear gene transformation in a dinoflagellate. doi: 10.1101/602821</cite></li>
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<li><cite>[3]  Ortiz-Matamoros, M.F., Islas-Flores, T., Voigt, B., Menzel, D., Baluška, F. & Villanueva, M.A. (2015, July 13). Heterologous DNA Uptake in Cultured
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Symbiodinium spp. Aided by Agrobacterium tumefaciens. doi:10.1371/journal. Pone.0132693
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</cite></li>
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GSU iGEM

Creating Synbio-dinium

Chasing Coral


Our Inspiration

The inspiration behind our project was the Netflix documentary Chasing Coral. From this documentary we learned about the extent of the devastation to coral reefs caused by bleaching. Many of the scientists seen in the film expressed despair over the seeming inevitability of the destruction. While current efforts to protect reefs have focused on enacting marine protected areas to prevent commercial fishing and industrial activities from harming the reefs, these efforts cannot stop the environmental effects of climate change on ocean waters. In the absence of a realistic proposal to halt global warming by eliminating fossil fuel consumption, marine heatwaves will become more frequent and even more severe ( Nature Communications volume 9, Article number: 1324 (2018), https://www.nature.com/articles/s41467-018-03732-9) leading to a corresponding increase in coral bleaching and reef loss. About half of the Great Barrier Reef died after the heat wave of 2016/2017.https://www.newscientist.com/article/2198318-the-great-barrier-reef-is-losing-its-ability-to-recover- from-bleaching/.And according to the UN Special Report on Global Warming, all global reefs around the world are expected to completely die off if global temperatures increase by another 2 degrees Celsius (https://www.ipcc.ch/2018/10/08/summary-for-policymakers-of-ipcc-special-report-on-global- warming-of-1-5c-approved-by-governments/).

The Problem

Coral bleaching, the loss of algal symbionts necessary for the survival of cnidarian reef organisms, is a disastrous environmental issue with global consequences. No single factor has been established as the cause of this catastrophe, but there are a multitude of suspects including increased greenhouse gas emissions and rising seawater temperatures[1].

The Solution

Although there is no absolute certainty about the cause of coral bleaching, increasing water temperatures have been directly correlated with subsequent bleaching events. And coral bleaching can be stimulated in aquarium corals by raising the water temperatures. There are some strains of corals and their symbiotic algae that prove somewhat more resistant to bleaching than others. So we proposed to identify those characteristics that might improve the ability to withstand bleaching and use a synthetic biology approach to modify the algae and restore them to corals.

The Plan

We are establishing both culturing and transformation protocols for these microalgae symbionts. We began by optimizing culturing techniques for Symbiodinium microadriaticum, Oxyrrhis marina (our model organism), and Dunaliella tertiolecta (the food source for O. marina).
To identify optimal algal growth conditions, we tested various factors such as growth media, light intensity, and temperature. We designed a codon optimized red fluorescent protein part that was cloned into a dinoflagellate-optimized expression plasmid (DinoIII)[2] for transformation into our model organism as a proof of concept. In parallel, we are also attempting to replicate the only known successful transformation of Symbiodinium using an Agrobacterium tumefacien co-culture carrying a binary vector, pCB302-GFP-MBD[3], and designing/executing various electroporation protocols. A genomic analysis of clade D Symbiodinium, a clade associated with higher resistance to bleaching but diminished coral growth, will identify target genes related to bleaching resistance for transformation into the growth-favorable clade C of Symbiodinium. The corals will then uptake the modified host algae, increasing their resistance to bleaching.
  • [1] Douglas (2003, April 4). Coral Bleaching--How and Why doi:10.1016/S0025-326X(03)00037-7/journal.
  • [2] Sprecher,B., Zhang,H. & Lin, S. (2019, April 9). Nuclear gene transformation in a dinoflagellate. doi: 10.1101/602821
  • [3] Ortiz-Matamoros, M.F., Islas-Flores, T., Voigt, B., Menzel, D., Baluška, F. & Villanueva, M.A. (2015, July 13). Heterologous DNA Uptake in Cultured Symbiodinium spp. Aided by Agrobacterium tumefaciens. doi:10.1371/journal. Pone.0132693
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    Tel: (+1) 404 413 5344

    E-mail: igemgsu@gmail.com

    Address: iGEM 50 Decatur St SE, Atlanta GA 30303