Team:Concordia/Project

From 2014.igem.org

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   <h2>Our Project Description</h2>
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   <h2>An introduction:</h2>
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  <p>Our team aims to engage in a synthetic biology project that carries significant relevance to current day problems. Following a thorough project exploration phase, our team decided on a project titled, “Clean Green Lipid Machine” which involves the genetic engineering of microalgae.</p>
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  <p>Microalgae are a varied group of organisms with great potential in synthetic biology. Compared to other iGEM &amp; genetic engineering model organisms, such as E. coli and yeast, microalgae distinguish themselves by the fact that they are photosynthetic with the potential of carbon neutral production of a variety of high valuable metabolites. These metabolites include biofuels, biolipids, proteins, hormones and antibodies, many of which are currently produced via resource intensive processes.</p>
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  <p>Unicellular microalgae are a varied group of organisms with excellent potential in applied and exploratory synthetic biology. Compared with the majority of other model and industrial organisms, most microalgae have the environmentally beneficial distinction of being able to carry out photosynthesis. With their photosynthetic and mixotrophic abilities, these organisms have the promise of becoming platforms for carbon-neutral production of both high value and inexpensive metabolites. Additionally, microalgae have the capability to express genes from plants, fungi and prokaryotes making them ideal hosts for combinatorial recombinant gene expression. </p>
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  <p>Our goal is to develop genetic engineering tools for microalgae, specifically various Chlorella &amp; Chlamydomonas strains , to turn them into a chassis for future iGEM teams and researchers. This will provide an organism capable of synthesizing a variety of useful molecules such as lipids, which can be used as biofuels, or modified into useful lipid based compounds like omega 3 fatty acids. </p>
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  <p>Non-engineered strains are already being used for biofuels and lipids. With the creation of standardised tools for the stable engineering of microalgae, the community will be able to continue asking deeper questions about basic biology and, drastically increase the promise of microalgae as an industrial chassis.</p>
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  <p>By building and characterising tools for genome scale engineering of multiple genera of microalgae, the Concordia iGEM team has opened a new world for the use of industrially relevant photosynthetic microbes by iGEM teams and researchers alike.
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   <h2>Our Plan of Action</h2>
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   <h2>The Concordia Microalgae Toolkit:</h2>
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  <p>One of our project goals is to create a library of useful and characterized parts; mainly by designing cassettes with varying promoters, genes and terminators from different species. These cassettes were constructed via Gibson assembly and then the plasmids containing the cassettes were inserted in Chlamydomonas and Chlorella via electroporation.</p>
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  <p>Our team grew out four different strains of Chlorella (Vulgaris, Ellipsodia, Saccharophilea, Kessleri) and a cell wall deficient strain of Chalmydomonas (UVM1). We characterized their growth patterns by obtaining various optical density and cell count measurements. With these results, we created growth curves and established the mid-log phase (ideal for transformation) of all of our strains.</p>
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  <p>Starting with the goal of making microalgae a viable platform in synthetic biology, we set out to create a toolkit of standardised biological parts for use in these organisms. We chose to work with a diverse group of species that included Chlorella vulgaris, Chlorella ellipsodia, Chlorella saccharophilea, Chlorella kessleri, and Chlamydomonas reinhardtii. These species present a broad range of optimum growth conditions and metabolic profiles. </p>
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  <p>We plan on using CrispR along with an NLS signal to direct the cas9 gene into the nucleus. By proving that our CrispR system works, we could safely assume that this system would work for the insertion of other genes of interest in Chlorella as well. </p>
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  <p>Characterised parts in our toolkit include promoters, terminators, fluorescent proteins, localisation tags, antibiotic markers, and CRISPR/Cas. </p>
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  <p>Our promoter library offers plant, fungal, and viral promoters of varying strengths for control of heterologous gene expression in microalgae. Our terminators were compared alongside a variety of these promoters. Multiple localization signals were investigated to enable a user to perform targeted gene expression. We also systematically characterised the effective concentration ranges for antibiotic use in a manner not yet seen for microalgae. </p>
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  <p>CRISPR/Cas systems have revolutionized genome engineering efforts by providing a quick and efficient means of inserting and deleting DNA from the host’s genome. Our toolkit set out to demonstrate the ability to perform targeted multi-plex genome engineering of microalgae in a manner accessible to any researcher.</p>
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Revision as of 20:42, 15 August 2014

iGEM Concordia 2014

An introduction:

Unicellular microalgae are a varied group of organisms with excellent potential in applied and exploratory synthetic biology. Compared with the majority of other model and industrial organisms, most microalgae have the environmentally beneficial distinction of being able to carry out photosynthesis. With their photosynthetic and mixotrophic abilities, these organisms have the promise of becoming platforms for carbon-neutral production of both high value and inexpensive metabolites. Additionally, microalgae have the capability to express genes from plants, fungi and prokaryotes making them ideal hosts for combinatorial recombinant gene expression.

Non-engineered strains are already being used for biofuels and lipids. With the creation of standardised tools for the stable engineering of microalgae, the community will be able to continue asking deeper questions about basic biology and, drastically increase the promise of microalgae as an industrial chassis.

By building and characterising tools for genome scale engineering of multiple genera of microalgae, the Concordia iGEM team has opened a new world for the use of industrially relevant photosynthetic microbes by iGEM teams and researchers alike.



The Concordia Microalgae Toolkit:

Starting with the goal of making microalgae a viable platform in synthetic biology, we set out to create a toolkit of standardised biological parts for use in these organisms. We chose to work with a diverse group of species that included Chlorella vulgaris, Chlorella ellipsodia, Chlorella saccharophilea, Chlorella kessleri, and Chlamydomonas reinhardtii. These species present a broad range of optimum growth conditions and metabolic profiles.

Characterised parts in our toolkit include promoters, terminators, fluorescent proteins, localisation tags, antibiotic markers, and CRISPR/Cas.

Our promoter library offers plant, fungal, and viral promoters of varying strengths for control of heterologous gene expression in microalgae. Our terminators were compared alongside a variety of these promoters. Multiple localization signals were investigated to enable a user to perform targeted gene expression. We also systematically characterised the effective concentration ranges for antibiotic use in a manner not yet seen for microalgae.

CRISPR/Cas systems have revolutionized genome engineering efforts by providing a quick and efficient means of inserting and deleting DNA from the host’s genome. Our toolkit set out to demonstrate the ability to perform targeted multi-plex genome engineering of microalgae in a manner accessible to any researcher.



Stay in touch with the iGEM Concordia 2014 team: