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| − | <h1>Our Project:</h1> | + | <div class="main-pg"> |
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| − | <p>Bacteria and yeast cells have been used for a long time in protein production, but their main problem is that they aren’t able to fold complex proteins correctly and to do some posttranslational modifications. Looking for a solution to this problem, we are developing a toolkit box for the use of Sf9 cells for protein production. Sf9 cells are derived from Spodoptera frugiperda, the Fall Armyworm Moth, and are currently used for the production of vaccines and antibodies. This project will introduce 3 promoters for the use in baculovirus transfection and 1 promoter for stable line generation, 2 secretion signal peptides, 2 reporters, and T2A, all of them to simplify the use of this insect cell platform. Additionally, we will produce the Epidermal Growth Factor Receptor. We will also contrast the protein production levels through baculovirus transfection and stable line generation; this will allow other teams to choose the protein production method that better suits their needs. To accomplish this, first of all, a standardization of the protocols needed for the Sf9 cells growth and transfection will be done. Then, the basic components for the construction of biological circuits will be characterized. During this step, two protein production methods are going to be used: baculovirus transfection and stable line generation. At last, the system will be validated with the production of a complex model protein with posttranslational modifications, Epidermal Growth Factor Receptor. By the inclusion of this platform, it is expected that its use in synthetic biology will be increased, due to its high similarities to mammalian cells at a lower cost. Even though we consider our project to be foundational, we have in mind the future use of the Sf9 cells as the primary organism in which biopharmaceuticals could be produced, as the maintenance costs could be reduced in comparison to mammalian cells.</p>
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| | + | <video style="width:100%" class="main-page" autoplay loop><source src="https://static.igem.org/mediawiki/2015/e/ee/Tec-Monterrey_Welcome_Video.mp4" type="video/mp4"></video> |
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| | + | <h1>Insects join iGEM: <i>Sf9</i> cells as a new chassis for synthetic biology</h1> |
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| | + | <h2>Project Description</h2> |
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| | + | The purpose of this project is to introduce the Sf9 cell line (Fall armyworm ovary, <i>Spodoptera frugiperda</i> cells) to synthetic biology and the iGEM Competition, by developing a tool kit of several biobricks for their use in this new chassis. We will use the Sf9 cells as an alternative for the production of proteins with high complexity and post-traductional modifications, because of their flexibility, cultivation time and minor cost compared with other expression systems. We will characterize functional parts for two main areas: Transfection and Genome Editing. For the first part we will characterize baculovirus mediated transfection and, direct plasmid transfection for the generation of stable cell lines; for the second part we will use the CRISPR/Cas9 system to assay highly specific genome modifications in an effort to improve production and ease of use of this technology. |
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