Difference between revisions of "Team:UCLA"

 
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<h2> Welcome to iGEM 2015! </h2>
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<p>Your team has been approved and you are ready to start the iGEM season! </p>
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<h1 style="position:relative;top:-20px;text-decoration:none;font-family: 'Nexa Light', sans-serif;color:white;" align="middle"><b>SilkyColi: Reprogramming the physical and functional properties of synthetic silks </b></h1>
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<h2> Project Descriptions (Due July 15th) </h2>
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<b> (very much a work in progress) Silky Coli: Exploring and designing unique modular parts for synthetic silk assembly and function. </b>
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<p> The use of silk fibroin produced by both spiders and silkworms have recently arisen as a suitable alternative to synthetic fiber products for a wide variety of cosmetic, engineering, and biomedical opportunities. However, efforts to develop high-throughput production of silk fibroin via an <i> in vivo </i> approach have proven to be unsustainable and highly costly to process. Work to produce recombinant fibroin using high expression <i> Escherichia coli </i> chassis has shown to be both effective and scalable, but the difficulty in generating a large cloning library with monomeric MaSP1/2 subunits for polymerization with various affinity binding domains prevents the creation of modular silk fibroins that can perform an array of functions. To add modular and scalable affinity domains to nascent silk fibroins, a composite silk construct containing the N- and C- terminal domains derived from <i> Nephila clavipes </i> spider silk constructions ligated to a reporting marker (initially, super folder GFP) flanked by Type IIS restriction sites was cloned and expressed in <i> E. coli </i> colonies. Preliminary results indicate high expression levels driven by an IPTG-inducible promoter hours after induction, and IMAC purification indicates high degree of protein yield and purity via electrophoretic analysis. Further characterization must be conducted to verify protein conformation and function using blotting analysis, spectroscopy, and fluorescence assaying. </p>
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<p>In our exploration of silk and silk materials, we came across honeybee silk from <i>Apis mellifera</i>. Although it may not have the tensile strength of spider silk, it is extremely flexible and has potential as a high performance material. It also has the advantage of being a non-repetitive sequence, which makes it much easier to work with genetically. We will submit the honeybee silk coding region as a novel biobrick to add to the registry's growing collection of protein material parts.  Furthermore, we plan on functionalizing the honeybee silk by first fusing it with GFP as a proof of principle, and later fusing it to affinity domains to create a modular fiber.  </p>
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<p>The third part of our project involves processing the recombinant silk proteins in order to create functional silk threads and documenting their characteristics. We are trying to do so with low-budget and easy-to-use protocols, so that future iGEM teams who pursue projects related to silk will be able to do so easily.</p>
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<h4>Before you start: </h4>
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<p> Please read the following pages:</p>
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<li> <a href="https://2015.igem.org/Requirements">Requirements page </a> </li>
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<li> <a href="https://2015.igem.org/Wiki_How-To">Wiki Requirements page</a></li>
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          <h1 align="middle">RESULTS</h1>
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        <center><img src="https://static.igem.org/mediawiki/2014/8/88/Goldmedal_header.png" width="120px"><h2>Gold Medal Award, Undergraduate Division</h2> <h2>Runner-Up for Best Manufacturing Project, Undergraduate Division</h2><h2>Top 5 for Best New Basic Part, Undergraduate Division</h2><h2>Top 5 for Best Part Collection, Undergraduate Division</h2></center>
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<h4> Styling your wiki </h4>
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<p>You may style this page as you like or you can simply leave the style as it is. You can easily keep the styling and edit the content of these default wiki pages with your project information and completely fulfill the requirement to document your project.</p>
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          <h1 align="middle">Abstract</h1>
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<p>While you may not win Best Wiki with this styling, your team is still eligible for all other awards. This default wiki meets the requirements, it improves navigability and ease of use for visitors, and you should not feel it is necessary to style beyond what has been provided.</p>  
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          <p>Among natural materials, silk fibers boast unparalleled strength and elasticity. This has made silk ideal for use in apparel, medical sutures, and other high-performance materials. The unique profile of silk arises from the composition of its repetitive protein domains, which varies between species. We aimed to program the physical properties of synthetic silk in two ways: by modularizing spider silk genes and tuning their properties through directed assembly, and by fusing accessory proteins to silkworm and honey bee silks to expand their functionality. To overcome the challenge of creating large, repetitive, GC-rich genes, we adapted Iterative Capped Assembly to ligate spider silk genes in specific ratios, orders, and lengths. The recombinant silks were expressed in E. coli then spun via standard wet spinning. This provides a platform for standardizing the customization of synthetic silk fibers, and exploring their potential as multipurpose biomaterials.</p>
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<h4> Editing your wiki </h4>
 
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<p>On this page you can document your project, introduce your team members, document your progress and share your iGEM experience with the rest of the world! </p>
 
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<p> <a href="https://2015.igem.org/wiki/index.php?title=Team:UCLA&action=edit"> Click here to edit this page! </a></p>
 
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<p>See tips on how to edit your wiki on the <a href="https://2015.igem.org/TemplatesforTeams_Code_Documentation">Template Documentation</a> page.</p>
 
  
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          <h1 align="middle" style="color:white">Projects</h1>
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<p> This year we have created templates for teams to use freely. More information on how to use and edit the templates can be found on the
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<a href="https://2015.igem.org/TemplatesforTeams_Code_Documentation">Template Documentation </a> page.</p>  
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    <h2 class="onBlack">Customizing Silk</h2>
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<h4>Tips</h4>
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Modifying the genetic structure of silk can create a diverse new range of biomaterials.
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<p>This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started: </p>
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<li>State your accomplishments! Tell people what you have achieved from the start. </li>
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We have adapted a novel cloning technique called iterative capped assembly (ICA) to rapidly and controllably assemble spider silk genes.  Our new process will dramatically shorten the time and expense needed to assemble these repetitive parts. Click here to learn more.
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<li>Be clear about what you are doing and how you plan to do this.</li>
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<li>You have a global audience! Consider the different backgrounds that your users come from.</li>
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<li>Make sure information is easy to find; nothing should be more than 3 clicks away.  </li>
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<li>Avoid using very small fonts and low contrast colors; information should be easy to read. </li>
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<li>Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the <a href="https://2015.igem.org/Calendar_of_Events">iGEM 2015 calendar</a> </li>
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<li>Have lots of fun! </li>
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  <a class="cover boxcaption" style="top: 340px;" href="https://2015.igem.org/Team:UCLA/Project/Functionalizing%20Silk">
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    <h2 class="onBlack">Functionalizing Fibers</h2>
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By attaching various proteins, we can create silk with specific applications.
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Glowing silk is an idea that appeals both to the scientific and artistic worlds. It is a proof of concept for the scientists, demonstrating how we are able to functionalize silk fibers with novel peptides, and an interesting new material to work with for the artists. This method can be applied to different kinds of proteins with different functions as well, but every idea has to start small. To see how we attached GFP to our silk constructs, click this text.
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<h4>Inspiration</h4>
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Engineering our silk is only half the work, creating a tangible product is the rest.
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<p> You can also view other team wikis for inspiration! Here are some examples:</p>
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<li> <a href="https://2014.igem.org/Team:SDU-Denmark/"> 2014 SDU Denmark </a> </li>
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We have developed a pipeline for processing and developing silk proteins produced into novel silks. Additionally, we have deigned methods to test the strength and elasticity of our fibers. To see the amazing fibers, films, and protocols we have created, click here to learn more!
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<li> <a href="https://2014.igem.org/Team:Aalto-Helsinki">2014 Aalto-Helsinki</a> </li>
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<li> <a href="https://2014.igem.org/Team:LMU-Munich">2014 LMU-Munich</a> </li>
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<li> <a href="https://2014.igem.org/Team:Michigan"> 2014 Michigan</a></li>
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<li> <a href="https://2014.igem.org/Team:ITESM-Guadalajara">2014 ITESM-Guadalajara </a></li>
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<li> <a href="https://2014.igem.org/Team:SCU-China"> 2014 SCU-China </a></li>
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<h4> Uploading pictures and files </h4>
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<p> You can upload your pictures and files to the iGEM 2015 server. Remember to keep all your pictures and files within your team's namespace or at least include your team's name in the file name. <br />
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When you upload, set the "Destination Filename" to <code>Team:YourOfficialTeamName/NameOfFile.jpg</code>. (If you don't do this, someone else might upload a different file with the same "Destination Filename", and your file would be erased!)</p>
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<a href="https://2015.igem.org/Special:Upload">CLICK HERE TO UPLOAD FILES</a>
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Latest revision as of 01:48, 11 October 2015

iGEM UCLA





SilkyColi: Reprogramming the physical and functional properties of synthetic silks


RESULTS

Gold Medal Award, Undergraduate Division

Runner-Up for Best Manufacturing Project, Undergraduate Division

Top 5 for Best New Basic Part, Undergraduate Division

Top 5 for Best Part Collection, Undergraduate Division

Abstract

Among natural materials, silk fibers boast unparalleled strength and elasticity. This has made silk ideal for use in apparel, medical sutures, and other high-performance materials. The unique profile of silk arises from the composition of its repetitive protein domains, which varies between species. We aimed to program the physical properties of synthetic silk in two ways: by modularizing spider silk genes and tuning their properties through directed assembly, and by fusing accessory proteins to silkworm and honey bee silks to expand their functionality. To overcome the challenge of creating large, repetitive, GC-rich genes, we adapted Iterative Capped Assembly to ligate spider silk genes in specific ratios, orders, and lengths. The recombinant silks were expressed in E. coli then spun via standard wet spinning. This provides a platform for standardizing the customization of synthetic silk fibers, and exploring their potential as multipurpose biomaterials.

Projects