Difference between revisions of "Team:XMU-China/Collaborations"

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     <title>Team:XMU-China/Description - 2018.igem.org</title>
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     <title>Team:XMU-China/Collaborations - 2018.igem.org</title>
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         <header>
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                                 <li><a href="https://2018.igem.org/Team:XMU-China/Description">Description</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Description">Description</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Design">Design</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Design">Design</a></li>
                                <li><a href="https://2018.igem.org/Team:XMU-China/Demonstrate">Demonstrate</a></li>
 
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Results">Results</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Results">Results</a></li>
 +
                                <li><a href="https://2018.igem.org/Team:XMU-China/Demonstrate">Demonstrate</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Parts">Parts</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Parts">Parts</a></li>
 
                             </ul>
 
                             </ul>
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                             <ul>
 
                             <ul>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Hardware">Overview</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Hardware">Overview</a></li>
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Hardware/Microfluidic_Chips">Microfluidic chips</a></li>
+
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Hardware#Microfluidic_Chips">Microfluidic Chips</a></li>
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Hardware/Fluorescenc_Detection">Fluorescence Detection</a></li>
+
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Hardware#Fluorescence_Detection">Fluorescence Detection</a></li>
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Hardware/Straberry_Pi">Straberry Pi</a></li>
+
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Hardware#Raspberry_Pi">Raspberry Pi</a></li>
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Applied_Design">Applied Design</a></li>
+
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Hardware#Application">Application</a></li>
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Software">APP</a></li>
+
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Software">Software</a></li>
 +
                                <li><a href="https://2018.igem.org/Team:XMU-China/Applied_Design">Product Design</a></li>
 
                             </ul>
 
                             </ul>
 
                         </li>
 
                         </li>
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                             <a href="#">Model</a>
 
                             <a href="#">Model</a>
 
                             <ul>
 
                             <ul>
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Model">Overview</a></li>
+
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Model#Summary">Summary</a></li>
 +
                                <li><a href="https://2018.igem.org/Team:XMU-China/Model#Thermodynamic_model">Thermodynamic Model</a></li>
 +
                                <li><a href="https://2018.igem.org/Team:XMU-China/Model#Fluid_dynamics_model">Fluid dynamics Model</a></li>
 +
                                <li><a href="https://2018.igem.org/Team:XMU-China/Model#Molecular_docking_model">Molecular Docking Model</a></li>
 +
                                <li><a href="https://2018.igem.org/Team:XMU-China/Model#The_dynamic_model">Derivation of Rate Equation</a></li>
 
                             </ul>
 
                             </ul>
 
                         </li>
 
                         </li>
 
                         <li class="Human_Practice">
 
                         <li class="Human_Practice">
                             <a href="#">Human Practice</a>
+
                             <a href="#">Social Works</a>
 
                             <ul>
 
                             <ul>
                                 <li><a href="https://2018.igem.org/Te
+
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Human_Practices">Human Practice</a></li>
                                am:XMU-China/Human_Practices">Overview</a></li>
+
                                <li><a href="https://2018.igem.org/Team:XMU-China/HP/Silver">Silver</a></li>
+
                                <li><a href="https://2018.igem.org/Team:XMU-China/HP/Gold_Integrated">Gold</a></li>
+
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Public_Engagement">Engagement</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Public_Engagement">Engagement</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Collaborations">Collaborations</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Collaborations">Collaborations</a></li>
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                                 <li><a href="https://2018.igem.org/Team:XMU-China/Notebook">Notebook</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Notebook">Notebook</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Experiments">Experiments</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Experiments">Experiments</a></li>
 +
                                <li><a href="https://2018.igem.org/Team:XMU-China/Engineering">Engineering</a></li>
 
                             </ul>
 
                             </ul>
 
                         </li>
 
                         </li>
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                                 <li><a href="https://2018.igem.org/Team:XMU-China/Attributions">Attributions</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Attributions">Attributions</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Judging">Judging</a></li>
 
                                 <li><a href="https://2018.igem.org/Team:XMU-China/Judging">Judging</a></li>
 +
                                <li><a href="https://2018.igem.org/Team:XMU-China/After_iGEM">After iGEM</a></li>
 
                             </ul>
 
                             </ul>
 
                         </li>
 
                         </li>
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         </header>
 
         </header>
 
     </div>
 
     </div>
    <script src="js/jquery-1.11.0.min.js"></script>
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     <script src="https://2018.igem.org/Team:XMU-China/js/hc-mobile-nav?action=raw&amp;ctype=text/javascript"></script>
    <!-- <script src="js/hc-mobile-nav.js"></script> -->
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     <script src="https://2018.igem.org/Team:XMU-China/js/hc-mobile-nav?action=raw&ctype=text/javascript"></script>
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     <div class="header">
 
     <div class="header">
 
         <div class="logo">
 
         <div class="logo">
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                         <li><a href="https://2018.igem.org/Team:XMU-China/Attributions">Attributions</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Attributions">Attributions</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Judging">Judging</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Judging">Judging</a></li>
 +
                        <li><a href="https://2018.igem.org/Team:XMU-China/After_iGEM">After iGEM</a></li>
 
                     </ul>
 
                     </ul>
 
                 </div>
 
                 </div>
 
                 <div id="Notebook">
 
                 <div id="Notebook">
                     <div>
+
                     <div class="nav-word">Notebook</div>
                        <div class="nav-word">Notebook</a></div>
+
                    </div>
+
 
                     <ul>
 
                     <ul>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Notebook">Notebook</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Notebook">Notebook</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Experiments">Experiments</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Experiments">Experiments</a></li>
 +
                        <li><a href="https://2018.igem.org/Team:XMU-China/Engineering">Engineering</a></li>
 
                     </ul>
 
                     </ul>
 
                 </div>
 
                 </div>
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                 </div>
 
                 </div>
 
                 <div id="Human_Practice">
 
                 <div id="Human_Practice">
                     <div class="nav-word">Human Practice</div>
+
                     <div class="nav-word">Social Works</div>
 
                     <ul>
 
                     <ul>
                         <li><a href="https://2018.igem.org/Team:XMU-China/Human_Practices">Overview</a></li>
+
                         <li><a href="https://2018.igem.org/Team:XMU-China/Human_Practices">Human Practice</a></li>
                        <li><a href="https://2018.igem.org/Team:XMU-China/HP/Silver">Silver</a></li>
+
                        <li><a href="https://2018.igem.org/Team:XMU-China/HP/Gold_Integrated">Gold</a></li>
+
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Public_Engagement">Engagement</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Public_Engagement">Engagement</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Collaborations">Collaborations</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Collaborations">Collaborations</a></li>
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                     <div class="nav-word">Model</div>
 
                     <div class="nav-word">Model</div>
 
                     <ul>
 
                     <ul>
                         <li><a href="https://2018.igem.org/Team:XMU-China/Model">Overview</a></li>
+
                         <li><a href="https://2018.igem.org/Team:XMU-China/Model">Summary</a></li>
 +
                        <li><a href="https://2018.igem.org/Team:XMU-China/Model#Thermodynamic_model">Thermodynamic Model</a></li>
 +
                        <li><a href="https://2018.igem.org/Team:XMU-China/Model#Fluid_dynamics_model">Fluid dynamics Model</a></li>
 +
                        <li><a href="https://2018.igem.org/Team:XMU-China/Model#Molecular_docking_model">Molecular Docking Model</a></li>
 +
                        <li><a href="https://2018.igem.org/Team:XMU-China/Model#The_dynamic_model">Derivation of Rate Equation</a></li>
 
                     </ul>
 
                     </ul>
 
                 </div>
 
                 </div>
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                     <ul>
 
                     <ul>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Hardware">Overview</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Hardware">Overview</a></li>
                         <li><a href="https://2018.igem.org/Team:XMU-China/Hardware/Microfluidic_Chips">Microfluidic chips</a></li>
+
                         <li><a href="https://2018.igem.org/Team:XMU-China/Hardware#Microfluidic_Chips">Microfluidic Chips</a></li>
                         <li><a href="https://2018.igem.org/Team:XMU-China/Hardware/Fluorescenc_Detection">Fluorescence Detection</a></li>
+
                         <li><a href="https://2018.igem.org/Team:XMU-China/Hardware#Fluorescence_Detection">Fluorescence Detection</a></li>
                         <li><a href="https://2018.igem.org/Team:XMU-China/Hardware/Straberry_Pi">Straberry Pi</a></li>
+
                         <li><a href="https://2018.igem.org/Team:XMU-China/Hardware#Raspberry_Pi">Raspberry Pi</a></li>
                         <li><a href="https://2018.igem.org/Team:XMU-China/Applied_Design">Applied Design</a></li>
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                         <li><a href="https://2018.igem.org/Team:XMU-China/Hardware#Application">Application</a></li>
                         <li><a href="https://2018.igem.org/Team:XMU-China/Software">APP</a></li>
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                         <li><a href="https://2018.igem.org/Team:XMU-China/Software">Software</a></li>
 +
                        <li><a href="https://2018.igem.org/Team:XMU-China/Applied_Design">Product Design</a></li>
 
                     </ul>
 
                     </ul>
 
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                         <li><a href="https://2018.igem.org/Team:XMU-China/Description">Description</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Description">Description</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Design">Design</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Design">Design</a></li>
                        <li><a href="https://2018.igem.org/Team:XMU-China/Demonstrate">Demonstrate</a></li>
 
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Results">Results</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Results">Results</a></li>
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                        <li><a href="https://2018.igem.org/Team:XMU-China/Demonstrate">Demonstrate</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Parts">Parts</a></li>
 
                         <li><a href="https://2018.igem.org/Team:XMU-China/Parts">Parts</a></li>
 
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             <div class="word">Description</div>
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             <div class="word">Collaborations</div>
 
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         </div>
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         <div class="Experiments hp">
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             <section id="Overview" class="js-scroll-step">
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                        <div class="col-md-offset-3 col-md-6">
                    <a href="#ABCDsystem"id="Quick_A">ABCDsystem</a></a>
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                <a href="#OMVs" class="Quick-navigation-item" >
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                                <div id="collapse0" class="panel-collapse collapse in" role="tabpanel">
                    <img id="turn_img" src="https://static.igem.org/mediawiki/2018/c/cd/T--XMU-China--right2.png">
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                                    <div class="panel-body">
                    <a href="#OMVs" id="Quick_B">OMVs</a></a>
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                                        <p> No one can make great progress without advice and golden ideas from their excellent partners. During last six month, we participated in some iGEM conferences or even worked as the innovative online meeting organizers. Hopefully these events contributed greatly to all of us.</p>
                <a href="#Supporting" class="Quick-navigation-item">
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                    <img id="turn_img" src="https://static.igem.org/mediawiki/2018/1/1c/T--XMU-China--right3.png">
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                                </div>
                    <a href="#Supporting" id="Quick_C">Supporting</a></a>
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                    ABCDsystem
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                 </div>
 
                 </div>
                <h1>Background</h1>
 
                <p>Protein plays a significant role in performing physiological functions<sup>[1]</sup>. However, in diseased cells, protein carrying out a certain function may indicate the proceedings of disease. Such protein could be sorted to biomarkers, which have been regarded as the targets of disease detection and treatment in recent years.<sup>[2]-[4]</sup> Therefore, detecting those biomarkers of protein-type becomes more and more critical to biological and medical fields.
 
                </p>
 
                <p>There are two main detecting approaches to detect a particular protein in a complex sample. One is direct determination of the content after purification, and the other is binding assays which include a target recognition probe and a signal transducer. The former approach includes gel filtration chromatography, ion exchange chromatography, nickel column and more. While on the down side, these methods involve high costs, strict equipment requirements and other drawbacks, which are not suitable for promotion and application. The enzyme-linked immunosorbent assay (ELISA) is a typical representative of the latter approach, nevertheless, such assays using antibodies as affinity ligands have cross-reactivity of antibodies compromising the specificity to the target of interest.<sup>[5]</sup> What’s worse, the premise of using ELISA is to find the corresponding antibodies, but the fact is that not all proteins can find their specific antibody protein. That is to say, the use of ELISA is also limited.
 
                </p>
 
                <p>In terms of binding assays, using aptamers as affinity ligands to recognize specific proteins are better than those using antibodies. Aptamers are short, synthetic single stranded oligonucleotides (DNA or RNA) that can bind to target molecules with high affinity and specificity.<sup>[6]-[9]</sup> They are commonly selected from random sequence libraries, using the systematic evolution of ligands by exponential enrichment (SELEX) techniques.<sup>[10]</sup> Advantages of aptamers over antibodies include longer shelflife, improved thermal stability and ease of modification and conjugation.<sup>[11]</sup>
 
                </p>
 
                <p>An interesting binding assay is to use aptamers as the target recognition probes and CRISPR-Cas12a (Cpf1) as the signal amplifier, which is called Aptamer Based Cell-free Detection system(ABCD system, Figure 1). We developed this system to detect those biomarkers of protein-type for the purpose of disease detection or staging.
 
                </p>
 
                <p class="F1">
 
                    <img src="https://static.igem.org/mediawiki/2018/e/e2/T--XMU-China--ABCD_system.png">
 
                    <p class="Figure_word">Figure 1. <strong>A</strong>ptamer <strong>B</strong>ased <strong>C</strong>ell-free <strong>D</strong>etection system.</p>
 
                </p>
 
                <h1>Abstract</h1>
 
                <p>The core of the ABCD system is the specific binding of the aptamer and its target protein. We immobilize the aptamer-“complementary strand” complex on a solid phase, using a “competitive” approach to free the “complementary strand”; then the “complementary strand” was detected using the trans-cleavage property of the Cpf1 protein, which allows the fluorescence recovery of the static quenched complex whose fluorophore and quencher are linked by a ssDNA. In summary, we initially transform the protein signal to the acid signal, then transform the nucleic acid signal to the fluorescence signal. We use aptamer SYL3C<sup>[12]</sup> against EpCAM, an epithelial cell adhesion molecule that is highly expressed on the surface of adenocarcinoma cells, to test the feasibility of our system.</p>
 
                <h1 class="reference">Reference</h1>
 
                <p>
 
                    [1] Janet Iwasa, Wallace Marshall. Karp's Cell and Molecular Biology: Concepts and Experiments (8th ed.). <i>Wiley: Hoboken, NJ.</i> <strong>2016</strong>, 48-49.
 
                    <br>[2] J. K. Aronson. Biomarkers and surrogate endpoints. <i>British Journal of Clinical Pharmacology.</i> <strong>2005</strong>, 59, 491-494.
 
                    <br>[3] Kyle Strimbu, Jorge A. Tavel. What are biomarkers? <i>Current Opinion in HIV and AIDS.</i> <strong>2010</strong>, 5, 463–466.
 
                    <br>[4] Biomarkers Definitions Working Group. Biomarkers and surrogate endpoints: Preferred definitions and conceptual framework. <i>Clin. Pharmacol. Ther.</i> <strong>2001</strong>, 69, 89-95.
 
                    <br>[5] Hongquan Zhang, Feng Li, Brittany Dever, Xing-Fang Li, X. Chris Le. DNA-Mediated Homogeneous Binding Assays for Nucleic Acids and Proteins. <i>Chem. Rev.</i> <strong>2013</strong>, 113, 2812-2841.
 
                    <br>[6] Larry Gold, Barry Polisky, Olke Uhlenbeck, Michael Yarus. Diversity of Oligonucleotide Functions. <i>Annu. Rev. Biochem.</i> <strong>1995</strong>, 64, 763-797.
 
                    <br>[7] Camille L.A. Hamula, Jeffrey W. Guthrie, Hongquan Zhang, Xing-Fang Li, X. Chris Le. Selection and analytical applications of aptamers. <i>Trends Anal. Chem.</i> <strong>2006</strong>, 25, 681-691.
 
                    <br>[8] Renee K. Mosing, Shaun D. Mendonsa, Michael T. Bowser. Capillary Electrophoresis-SELEX Selection of Aptamers with Affinity for HIV-1 Reverse Transcriptase. <i>Anal. Chem.</i> <strong>2005</strong>, 77, 6107-6112.
 
                    <br>[9] Maxim Berezovski, Andrei Drabovich, Svetlana M. Krylova, Michael Musheev, Victor Okhonin, Alexander Petrov, Sergey N. Krylov. Nonequilibrium Capillary Electrophoresis of Equilibrium Mixtures: A Universal Tool for Development of Aptamers. <i>J. Am. Chem. Soc.</i> <strong>2005</strong>, 127, 3165-3171.
 
                    <br>[10] M Darmostuk, S Rimpelova, H Gbelcova, T Ruml. Current approaches in SELEX: an update to aptamer selection technology. <i>Biotechnology Advances.</i> <strong>2015</strong>, 33, 1141-1161.
 
                    <br>[11] Sumedha D. Jayasena. Aptamers: an emerging class of molecules that rival antibodies in diagnostics. <i>Clin. Chem.</i> <strong>1999</strong>, 45, 1628-1650.
 
                    <br>[12] Yanling Song, Zhi Zhu, Yuan An, Weiting Zhang, Huimin Zhang, Dan Liu, Chundong Yu, Wei Duan, Chaoyong James Yang. Selection of DNA Aptamers against Epithelial Cell Adhesion Molecule for Cancer Cell Imaging and Circulating Tumor Cell Capture. <i>Anal Chem.</i> <strong>2013</strong>, 85, 4141-4149.
 
                </p>
 
 
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             </section>
             <section id="OMVs" class="js-scroll-step">
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             <section id="Competition_experiment" class="js-scroll-step">
                 <div class="headline">
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                 <div class="container">
                     OMVs
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                     <div class="row">
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                        <div id="accordion">
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                                <div class="panel-heading" role="tab" id="heading114-1">
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                                    The CCiC
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                                </a>
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                                        <p> The 5<sup>th</sup> CCiC , which full name is the Conference of China iGEMer Community Meetup, was held at ShanhaiTech University on the subject of “Among us, past, present and future meeting” from August 27<sup>th</sup> to 31<sup>th</sup>. Dozens of teams from all over the China as well as the Executive Vice President and Chief Operating Officer if the iGEM Foundation, Meagan Lizarazo took part in. During this conference, we fortunately got the chance to have deep communication with many iGEM teams including iGEM_BIT(Beijing Institute of Technology University), iGEM Tianjin (Tianjin Unniversity) and LZU_China (Lanzhou University), and here we really appreciated their advices on our project about the competitive experiments, Kai ABC systems and the OMVs systems. Luckily, we won the best presentation award. What's more, we were greatly benefited from Meagan’s lecture pointing out the importance of “After iGEM”. This strengthened our determination to contact the old members of XMU-China and luckily we got precious experiences and further understanding about this competition.</p>
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                                        <p class="F3"><img src="https://static.igem.org/mediawiki/2018/f/f6/T--XMU-China--hp-part4-1.jpg"></p>
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                                            <p class="F25"><img src="https://static.igem.org/mediawiki/2018/e/e5/T--XMU-China--hp-part4-3.jpg"></p>
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                                    The 6<sup>th</sup> Asia-Pacific iGEM Conference
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                                        <p>From July 30<sup>th</sup> to August 3<sup>rd</sup>, 2018, we participated in the 6th Asia-Pacific iGEM Conference hosted by National Chung Hsing University, Taichung. There was a total of 22 teams and 140 participants from Taiwan, China, and Japan, warming up for the final Jamboree in Boston at the end of October. </p>
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                                        <p class="F3"><img src="https://static.igem.org/mediawiki/2018/1/1a/T--XMU-China--team_banner2.png"><img src="https://static.igem.org/mediawiki/2018/1/13/T--XMU-China--collaboration-1.jpg"></p>
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                                        <p>The conference invited wonderful speakers from the field of Synthetic Biology and Ambassador of the iGEM HQ to give a talk at the Opening Ceremony. Among them, Prof. Kenji Tsuge from Kobe University explained the emerging synthetic biology technology that can link quantities of large fragments of genes and perform these genes; Dr. Zhang Jui-Jen, an associate researcher at China Medical University Hospital, shared his experience in using synthetic biology entrepreneurship; iGEM Asian Ambassador Chen Hong explained some new systems and shared experience on behalf of iGEM. We gained tons of new ideas from these lectures, which had great influences on our next work.</p>
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                                        <p class="F3"><img src="https://static.igem.org/mediawiki/2018/d/d8/T--XMU-China--collaboration-2.jpg"></p>
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                                        <p>In the five-day conference, we also had a 20-minute English presentation and a poster exhibition. It was the first time to present our project frankly to iGEMers from Asia, and a good opportunity to practice for the final Jamboree. At the end of the exhibition, professor Yang Wen-Ming from the field of molecular biology at National Chung Hsing University gave a comment on our presentation. Through the conference, we also made suggestions to other Asian teams, longing for more cooperation opportunities.</p>
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                                        <p>This conference has provided us tremendous new ideas to improve our project and final presentation.</p>
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                                        <p class="F3"><img src="https://static.igem.org/mediawiki/2018/6/62/T--XMU-China--hp-part4-4.jpg"></p>
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                                    Collaboration with SUSTech
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                                        <p class="F25"><img src="https://static.igem.org/mediawiki/2018/d/d3/T--XMU-China--collaboration-3.jpg">
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                                            <p class="Figure_word">SUSTech iGEM</p>
 +
                                        </p>
 +
                                        <p>This year, the SUSTech iGEM team used the microfluidic chip as the core carrier to build a complete hardware platform, which was very similar to the idea of XMU-China team in hardware design. On this basis, these two teams had an online communication on issues related to microfluidic chips.</p>
 +
                                        <p>SUSTech used high-precision and single-channel chip tubes to achieve cell sorting functions, with a peristaltic pump that provides driving force, and a flow meter that filters and encapsulates cell droplets. These devices greatly improve the precision of the entire device.</p>
 +
                                        <p>XMU-China also made a relevant introduction about the design of microfluidic chips. What's more, our team additionally answered questions about how to control liquid flow in the centrifugal force-driven microfluidic chip.</p>
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                                        <p class="F7"><img src="https://static.igem.org/mediawiki/2018/9/99/T--XMU-China--collaboration-4.jpg"><img src="https://static.igem.org/mediawiki/2018/c/c5/T--XMU-China--collaboration-5.jpg">
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                                            <p class="Figure_word"><strong>Figure 1</strong>:The design of microfluidic chips of SUSTech iGEM(left) and XMU-China(right).</p>
 +
                                        </p>
 +
                                        <p>In terms of the project, we believe SUSTech iGEM team's using chips to study the interaction between cells is very innovative. From our professional perspective, we gave some advice on cell signaling pathways and cell-cell interactions, and recommended several related articles to them, hoping to help them. Based on our project, they raised some questions about whether our cell-free system belongs to the field of synthetic biology. We put forward our views, and the two parties had a futher discussion. We believe that synthetic biology is not a simple use of biobrick to construct gene loops. Instead, it should be a broader definition. We think that standard components are like building blocks, but it is up to people to decide what items should be joint together, rather than building blocks themselves. Therefore, biobrick can be used to build systems outside the genetic loop.</p>
 +
                                        <p>In addition, both of us exchanged ideas on the Art&Design. We carried out visual design communications with SUStech on aspects involving poster, Wiki and PPT. We showed them our methods and ideas in visual designing, including drawing graphics of experimental process and other details.</p>
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                                    Collaboration with BIT
 +
                                </a>
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                                        <p>At CCiC, we are very delighted to find that BIT projects have a lot of intersections with us. We have similar representations.Therefore, our team communicated with them in the following three aspects.</p>
 +
                                        <p> The first is the exchange of ideas on the subject. We have had a two-hour telephone conversation with them. On the phone, we exchanged the purpose of the project and some detailed designs, introduced our own project to them and proposed some questions for each other.</p>
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                                        <p class="F7"><img src="https://static.igem.org/mediawiki/2018/5/51/T--XMU-China--collaboration-6.jpg"><img src="https://static.igem.org/mediawiki/2018/e/ee/T--XMU-China--collaboration-7.jpg"></p>
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                                            <p>Secondly, we asked the BIT team about a difficulty in the experimental program. I am very happy that they are effective. We proved that our aptamers and complementary chains are combined. We gave the results back to them and gave them suggestions of improvement. In addition, we also exchanged some experimental methods and protocol.</p>
 +
                                            <p>Finally, in order to prove the feasibility of their respective experimental designs, we mailed some materials to each other and successfully verified the effectiveness of the other's combination, which is also a very effective part of our cooperation.</p>
 +
                                            <p>The experimental results are shown in the figure below. Well 1: Aptamers, Well 2: Complementary chains, Well 3: Aptamers + Complementary chains. As can be seen from the figure, the electrophoresis speed of the combined DNA is slower than that of the aptamer and the complementary strand, and it is indeed combined.</p>
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                                    </div>
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                                    Collaborations-NAU-CHINA
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                                        <p>
 +
                                            On July 31, 2018, at the 6<sup>th</sup> Asia-Pacific iGEM Conference hosted by National Chung Hsing University, Taichung,we met with NAU-CHINA team and established a deep friendship. Since then, our two teams have exchanged and discussed "how to build a team." <p>The professor affirmed the feasibility of our project and gave us some invaluable advice. Under the suggestion of Professor Zhu Zhi, we chose EpCAM and aptamer SYL3C. In addition, we are very grateful to Professor Zhu for her support on many instruments and materials provided during our experiments.</p>
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                                            <p class="F3"><img src="https://static.igem.org/mediawiki/2018/5/52/T--XMU-China--collaboration-8.jpg"></p>
 +
                                                <p>First we discussed "what kind of team should we build?" The International Genetic Engineering Machine Competition is a creative synthetic biology event and an interdisciplinary competition involving cross-disciplinary fields in mathematics, computers, statistics and so on. So our team should be a creative, comprehensive synthetic biology team. Our team members should be creative and comprehensive students. Most students of our two teams have multiple roles in the team.</p>
 +
                                                <p>Second, we discussed "how to find new partners." After we know what kind of team we want to build, we could find the companions we need. First, we have presentations, exhibitions, lab activities, etc. to make students realize that, what kind of the team they will join in and what is the team building for? what role they will be in the team, and what should be done by themselves? In order to join this team and make this team better, how should them prepare and learn? Then we divide the interviewers into different groups according to different division of labor and their wishes. Different groups determine the way of assessment according to the division of labor. For example, the number module is tested by some models, and the art team performs preliminary selection through some art production tasks... We will give priority to students with multiple skills.</p>
 +
                                                <p>Third, we communicated on how to train the new team members in the early stages of team formation. First of all, we will introduce our team and International Genetic Engineering Machine Competition through presentations, posters, WeChat pushes, etc., and also convey our requests for new recruits. We will then train the interviewees in groups and we will select them during the training process. Our training is mainly to share the learning materials for students and let them learn by themselves.</p>
 +
                                                <p>Our two teams made recommendations on each other for training new members. They suggested us add a literature reading part to the training to teach students how to read literatures and how to extract important information from the literature. Then they shared with us some materials for teaching students to read the literature. Their advice has helped us a lot, and we plan to adopt their suggestions to improve this year's training. At the same time, we also suggest that they increase the opportunities for students to practice, let the students design their own topics, increase their free space, make them more creative and have more new ideas. In addition, we exchanged our training materials on modeling, hardware, software, art and web design with each other.</p>
 +
                                                <p>Finally, we communicated with the daily management of the team. At the beginning of each team, all the students will participate in the design of the project and conduct laboratory training. After the preliminary determination of the subject, the students were divided into different groups, responsible for different labors like experiments, modeling, human practice and so on. We hold group meetings every week, each group reports the progress of the project, and different groups communicate with each other. In addition, we will also hold regular entertainment activities such as picnics, parties, etc. to relax and enhance team friendship. Both of our teams have one thing in common. Most members are in multiple roles, which better strengthens the communication between groups. It shows that iGEM competition is a common combination. There are different groups in this team. Everyone is not working separately, but is always intimately connected.</p>
 +
                                                <p>Through the exchanges with the NAU-iGEM team, Our team has a better construction model, and we have more experience in the follow-up training this year. Thanks to the NAU team for their help.</p>
 +
                                    </div>
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                                    Collaborations with H14Z1_Hangzhou
 +
                                </a>
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                                        <p>
 +
                                            This year XMU-china team and H14Z1_hangzhou team's cooperation can be divided into the following two parts: </p>
 +
                                        <p>First, XMU-China team helped and instructed H14Z1_hangzhou team to fill out the judging form, and to offer some advice about the relevant representations of one of a previous year biobricks to meet the bronze standard four. In addition, we also assisted H14Z1_hangzhou to complete the design of the first level menu and the second level menu in the navigation bar, so that their results can be better displayed in front of the judges. </p>
 +
                                        <p class="F3"><img src="https://static.igem.org/mediawiki/2018/9/9d/T--XMU-China--collaboration-9.jpg"></p>
 +
                                            <p>Second, XMU-China and H14Z1_hangzhou assumed the task of the experimental representation each other. H14Z1_hangzhou helped XMU-China measure the fluorescence characterization data of BBa_K2623025. XMU-China helped H14Z1_hangzhou complete the experimental curve of the number of engineering bacteria in the process of fermenting yoghurt with the change of time.</p>
 +
                                            <p>The experimental steps and results are as following:</p>
 +
                                            <p>Experimental steps: </p>
 +
                                            <p>1. The first day, Lactobacillus NZ9000 and GMCA respectively were inoculated into 50ml GM17 medium, fermented at 30 ℃ overnight (about 12h). (Place them in a 30 ℃ incubator, no shaker shocks required). </p>
 +
                                            <p>2. Next day, fresh milk was heated to about 55-60 degrees Celsius, and then adding some white sugar, 3000rpm stirring for 10 minutes mixing. Divided them into 250ml shaker, and the volume of liquid in every shaker is 100ml. </p>
 +
                                            <p>3. Sterilize the above solution at 90 ℃ for 5min, then cool it to 30 degrees Celsius to prepare for inoculation and fermentation. Then collect Lactobacillus acidophilus in shaker and clean them 1 times with sterile water. Add Lactobacillus acidophilus to the sample making the initial OD is 0.1 (600nm). Then let them ferment statically in the 250ml shaker at 30 ℃ (Place them in the 30 ℃ incubator, without shaker vibration) </p>
 +
                                            <p>4. Measure the OD value every two hours and count the corresponding number of cells to draw the curve. Simultaneously, test the pH of the solution (Measure for 14 hours)</p>
 +
                                            <p>Experimental results:</p>
 +
                                            <p>Table 1 The comparison of biomass and product formation in two different yogurt</p>
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                                            <div class="table_container">
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                                                <table class="table">
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                                                    <tr>
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                                                        <td></td>
 +
                                                        <td colspan="2">Cell number ( /ml)</td>
 +
                                                        <td colspan="2">GSH (mg/L)</td>
 +
                                                        <td colspan="2">SAM (mg/L)</td>
 +
                                                       
 +
                                                       
 +
                                                    </tr>
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                                                    <tr>
 +
                                                        <td>Strain</td>
 +
                                                        <td>6h</td>
 +
                                                        <td>12h</td>
 +
                                                        <td>6h</td>
 +
                                                        <td>12h</td>
 +
                                                        <td>6h</td>
 +
                                                        <td>12h</td>
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                                                    </tr>
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                                                    <tr>
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                                                        <td>L. lactis NZ9000
 +
                                                        </td>
 +
                                                        <td>3.5x 107</td>
 +
                                                        <td>1.1 x 109</td>
 +
                                                        <td>0.0</td>
 +
                                                        <td>0.0</td>
 +
                                                        <td>0.3</td>
 +
                                                        <td>0.8</td>
 +
                                                    </tr>
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                                                    <tr>
 +
                                                        <td>L. lactis NZ9000/pGMcA</td>
 +
                                                        <td>3.1x 107</td>
 +
                                                        <td>1.0 x 109</td>
 +
                                                        <td>4.5</td>
 +
                                                        <td>11.8</td>
 +
                                                        <td>1.5</td>
 +
                                                        <td>2.6</td>
 +
                                                    </tr>
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                                                </table>
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                                    Collaborations with FJNU-China
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                                </a>
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                                        <p>
 +
                                            2018 is the first year for FJNU-China team to participate in the iGEM competition, yet the teams competing in the first year are often lack of the relevant experience and grasp of the subject scale.</p>
 +
                                            <p>Since XMU-China team and FJNU-China are the only two teams in Fujian province in China, it is incumbent upon XMU-China team to instruct FJNU-China team and communicate with them to make the team fell they are parts of iGEM family as soon as possible.</p>
 +
                                            <p class="F3"><img src="https://static.igem.org/mediawiki/2018/3/3c/T--XMU-China--logo_pink.png"><span class="F7"><img src="https://static.igem.org/mediawiki/2018/2/2f/T--XMU-China--collaboration-10.jpg"></span></p>
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                                            <p>In August, we held the Conference of Fujian Province iGEMer Community in China,and at the meeting, the both teams presented their own topics. Based on the topic of ideas, gene circuit design, experimental programs and practical application level, XMU-China put forward their own ideas, which helped FJNU-China better prepare for the competition and explain the ideas of their subject. After the meeting, we introduced them to the hospital's dermatologist and led them to visit the Xiamen Environmental Protection Bureau.</p>
 +
                                            <p class="F3"><img src="https://static.igem.org/mediawiki/2018/e/e6/T--XMU-China--collaboration-11.jpg"></p>
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                                            <p>At the end of the competition, XMU-China provided years of experience to help FJNU-China to complete the safety form and judging form.</p>
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                                    SYSU-Software
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                                        <p>In the brainstorming stage, in order to solve the problem of preservation of Cas12a protein in microfluidic chips in hardware design, our team retrieved a large number of topics related to protein preservation in previous years. Finally inspired by 2017 TU_Delft's TDPs project and improving the characterization method. But it costed a lot of our time and efforts. After communicating with the team SYSU_Software in September, we asked them to quickly and accurately search for the previous project. They upgraded the functions of their software to  cater our needs. We compared the information obtained by the manual search and the information obtained through the software search, and found that the software can be well positioned to the part of the experimental plan we need, which is equivalent to the accuracy of the manual search, but the work efficiency is improved and the work efficiency is lowered. After obtaining the results, we made recommendations on the user-friendliness of the software's user interface operations and the search algorithms can be optimized later.
 +
                                        </p>
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                                        <p class="F7"><img src="https://static.igem.org/mediawiki/2018/c/c2/T--XMU-China--collaboration-12.jpg">
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                                            &nbsp<span class="F3"><img src="https://static.igem.org/mediawiki/2018/6/6c/T--XMU-China--collaboration-13.jpg"></span>
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                                        </p>
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                                    Tianjin iGEM
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                                        <p>This year, the Tianjin team in China worked on Clock Mechanisms, which would utilize some circadian clock proteins discovered in Synechococcus elongatus (strain PCC 7942) (Anacystis nidulans R2).</p>
 +
                                        <p>We XMU-China also made efforts in Circadian clock “Kai”. Unfortunately, three kinds of protein which we used can not be produced in our lab, because the DNA ordered from IDT was disabled.</p>
 +
                                        <p class="F25"><img src="https://static.igem.org/mediawiki/2018/1/11/T--XMU-China--collaboration-14.jpg"></p>
 +
                                        <p>However, when we talked about circadian clock with members in Tianjin Team, we were lucky enough to be informed that they could provide us with the DNA we needed. Thanks to their donations, our study on rebuilding circadian clock “Kai” in E.coli can proceed.</p>
 +
                                        <p>More details about Tianjin’s work can be found in https://2018.igem.org/Team:Tianjin</p>
 +
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                                    Newsletter
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                                        <p>This year we have newsletters with other iGEM teams from schools of all over the world, like Boston University (BU), Technische Universiteit Delft(TUDelft),TUST, ECUST, JNU and so on. Those letters are precious and beneficial since they enable different teams to share projects and views with each other, especially on topics like team information, team introduction, project description, etc. There is a link that you can see the new version of <a class="downloadpdf" href="https://static.igem.org/mediawiki/2018/0/0b/T--XMU-China--newsletter.pdf">the newsletters</a>. Through this platform, more people could learn about our projects and we have more opportunities to communicate and cooperate with the external world.
 +
                                        </p>
 +
                                       
 +
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 +
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 +
                         
 +
                        </div>
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                    </div>
 
                 </div>
 
                 </div>
                <h1>Background</h1>
 
                <p>Outer-membrane vesicles (OMVs) are lipid vesicles commonly produced by Gram-negative bacteria, which are filled with periplasmic content and are 20-250 nm in diameters (Figure 1). The production of OMVs allows bacteria to interact with their environment, and OMVs have been found to mediate diverse functions, including promoting pathogenesis, and enabling bacterial delivery of nucleic acids and proteins. A recent paper by Kojima R et al. 2018, demonstrated an EXOtic device that can produce exosomes with specific nucleic acids cargo (Figure 2). We were impressed by the amazing OMVs and EXOtic device and came up with an idea to design a cell-free system to enable specific siRNA to be encapsulated into OMVs for cancer treatment.
 
                </p>
 
                <p class="F2">
 
                    <img src="https://static.igem.org/mediawiki/2018/4/43/T--XMU-China--OMVs11.png">
 
                    <p class="Figure_word">Figure 2. The cell envelope of Gram-negative bacteria consists of two membranes, the outer membrane and the cytoplasmic membrane. Envelope stability comes from various crosslinks including the non-covalent interactions between the PG and the porin outer-membrane protein A (OmpA).</p>
 
                </p>
 
                <p class="F2">
 
                    <img src="https://static.igem.org/mediawiki/2018/c/c0/T--XMU-China--OMVs12.png">
 
                    <p class="Figure_word">Figure 3. Schematic illustration of the EXOtic devices. Exosomes are nanoscale extracellular lipid bilayer vesicles of endocytic origin, and they are secreted by nearly all cell types in physiological and pathological conditions. Exosomes containing the RNA packaging device (CD63-L7Ae) and mRNA (e.g., nluc-C/Dbox) can efficiently deliver specific nucleic acids.</p>
 
                </p>
 
                <h1>Abstract</h1>
 
                <p>Not only eukaryotes but also prokaryotes can produce nanoscale bubbles to fulfill diverse functions, such as cellular communication, surface modifications and the elimination of undesired components. Additionally, because of this functional versatility, OMVs have been explored as a platform for bioengineering applications. This year, we XMU-China decide to utilize OMVs as a cell-free platform to deliver our nucleic acids agents to facilitate therapeutic targeting of oncogenic KRAS in pancreatic cancer.</p>
 
                <p class="F3">
 
                    <img src="https://static.igem.org/mediawiki/2018/9/9d/T--XMU-China--OMVs13.png">
 
                    <p class="Figure_word">Figure 4. We utilize a split protein SpyTag/SpyCatcher (ST/SC) bioconjugation system to create a synthetic linkage between protein OmpA and archaeal ribosomal protein L7Ae. We fuse SpyTag with OmpA at its C-termini and N-termini respectively.</p>
 
                </p>
 
                <p class="F3">
 
                    <img src="https://static.igem.org/mediawiki/2018/d/da/T--XMU-China--OMVs14.png">
 
                    <p class="Figure_word">Figure 5. After the induction of IPTG and Arabinose, we can get L7Ae-SpyCatcher and siRNA-C/Dbox. Archaeal ribosomal protein L7Ae owns the ability to bind with C/Dbox RNA structure.</p>
 
                </p>
 
                <p class="F4">
 
                    <img src="https://static.igem.org/mediawiki/2018/9/97/T--XMU-China--OMVs15.png">
 
                    <p class="Figure_word">Figure 6. With the interaction between SpyTag and SpyCatcher, and the ability of L7Ae to be bind with C/Dbox, we can produce customizable and cell-free OMVs containing specific siRNA to traget for oncogenic gene.</p>
 
                </p>
 
                <h1 class="reference">Reference</h1>
 
                <p>
 
                    [1] Kojima R, Bojar D, Rizzi G, et al. Designer exosomes produced by implanted cells intracerebrally deliver therapeutic cargo for Parkinson’s disease treatment[J]. <i>Nature Communications.</i> <strong>2018</strong>, 9(1):1305. <br>
 
                    [2] Alves N J, Turner K B, Medintz I L, et al. Protecting enzymatic function through directed packaging into bacterial outer membrane vesicles: [J]. <i>Scientific Reports</i>, <strong>2016</strong>, 6:24866. <br>
 
                    [3] Schwechheimer C, Kuehn M J. Outer-membrane vesicles from Gram-negative bacteria: biogenesis and functions. [J]. <i>Nature Reviews Microbiology</i>, <strong>2015</strong>, 13(10):605-19. <br>
 
                    [4] Vanaja S K, Russo A J, Behl B, et al. Bacterial Outer Membrane Vesicles Mediate Cytosolic Localization of LPS and Caspase-11 Activation. [J]. <i>Cell</i>, <strong>2016</strong>, 165(5):1106-1119. <br>
 
                    [5] Kamerkar S, Lebleu V S, Sugimoto H, et al. Exosomes facilitate therapeutic targeting of oncogenic KRAS in pancreatic cancer[J]. <i>Nature</i>, <strong>2017</strong>, 546(7659):498-503. <br>
 
                    [6] https://en.wikipedia.org/wiki/Pancreatic_cancer<br>
 
                </p>
 
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                <div class="headline">
 
                    Supporting
 
                </div>
 
                <h1>Background</h1>
 
                <p>Tardigrades are able to tolerate almost complete dehydration by reversibly switching to an ametabolic state. This ability is called anhydrobiosis.<sup>[1]</sup>Tardigrade-specific intrinsically disordered proteins (TDPs) are essential for desiccation olerance.<sup>[2]</sup>2012, Takekazu Kunieda and his team identified five abundant heat-soluble proteins in the tardigrades, which can prevent protein-aggregation in dehydrated conditions in other anhydrobiotic organisms.<sup>[1]</sup>
 
                </p>
 
                <p class="F4">
 
                    <img src="https://static.igem.org/mediawiki/2018/2/2d/T--XMU-China--TDP1.png">
 
                    <p class="Figure_word">Figure 7. Stage Photo of Tardigrades in Ant-Man 2.</p>
 
                </p>
 
                <p>In 2017, Thomas C. Boothby and his team segregated three TDP proteins in the water bears and explored their mechanism of action<sup>[3]</sup>. This is a schematic diagram of the mechanism they have done so far. At the same time, one of the 2017 iGEM teams <a href="https://2017.igem.org/Team:TUDelft/Design"><span class="click_here">TUDelft</span></a>, attempted to preserve the Cas13a protein using the TDP proteins, and they also tried to preserve the bacteria with the TDP proteins and obtained amazing outcome.
 
                    In our project, we are going to use TDPs to help preserve the protein Cas12a and OMVs.
 
                </p>
 
                <h1>Abstract</h1>
 
                <p>We have carried out research on TDP proteins this year. On the one hand, we plan to preserve the Cas12a required for protein detection and OMVs required for treatment with TDPs. On the other hand, as the wiki says, TDP is a new biological activity protector with great potential. So we are going to use TDP proteins to simplify existing methods of preserving proteins and bacteria.
 
                    There are two novel protein families with distinct subcellular localizations named Cytoplasmic Abundant Heat Soluble (CAHS) and Secretory Abundant Heat Soluble (SAHS) protein families, according to their localization. In our project, SAHS1 was used to preserve the proteins and CAHS1 was used for the preservation of the bacteria.
 
                </p>
 
                <p class="F4">
 
                    <img src="https://static.igem.org/mediawiki/2018/a/aa/T--XMU-China--TDP2.png">
 
                    <p class="Figure_word">Figure 8. The Expression of TDPs When The Tardigrades Suffer Form Fast Drying and Slow Drying.(Thomas C. Boothby et al. 2017).</p>
 
                </p>
 
                <h1>Reference</h1>
 
                <p class="reference">
 
                    [1]. Yamaguchi A. Two Novel Heat-Soluble Protein Families Abundantly Expressed in an Anhydrobiotic Tardigrade. <i>PLoS ONE</i>, <strong>2012</strong>;7(8):e44209. <br>
 
[2]. Boothby TC. Tardigrades Use Intrinsically Disordered Proteins to Survive Desiccation. <i>Mol Cell</i>. <strong>2017</strong> Mar16;65(6):975-984.e5.
 
 
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Revision as of 21:55, 17 October 2018

Team:XMU-China/Collaborations - 2018.igem.org

Collaborations

No one can make great progress without advice and golden ideas from their excellent partners. During last six month, we participated in some iGEM conferences or even worked as the innovative online meeting organizers. Hopefully these events contributed greatly to all of us.

The 5th CCiC , which full name is the Conference of China iGEMer Community Meetup, was held at ShanhaiTech University on the subject of “Among us, past, present and future meeting” from August 27th to 31th. Dozens of teams from all over the China as well as the Executive Vice President and Chief Operating Officer if the iGEM Foundation, Meagan Lizarazo took part in. During this conference, we fortunately got the chance to have deep communication with many iGEM teams including iGEM_BIT(Beijing Institute of Technology University), iGEM Tianjin (Tianjin Unniversity) and LZU_China (Lanzhou University), and here we really appreciated their advices on our project about the competitive experiments, Kai ABC systems and the OMVs systems. Luckily, we won the best presentation award. What's more, we were greatly benefited from Meagan’s lecture pointing out the importance of “After iGEM”. This strengthened our determination to contact the old members of XMU-China and luckily we got precious experiences and further understanding about this competition.

From July 30th to August 3rd, 2018, we participated in the 6th Asia-Pacific iGEM Conference hosted by National Chung Hsing University, Taichung. There was a total of 22 teams and 140 participants from Taiwan, China, and Japan, warming up for the final Jamboree in Boston at the end of October.

The conference invited wonderful speakers from the field of Synthetic Biology and Ambassador of the iGEM HQ to give a talk at the Opening Ceremony. Among them, Prof. Kenji Tsuge from Kobe University explained the emerging synthetic biology technology that can link quantities of large fragments of genes and perform these genes; Dr. Zhang Jui-Jen, an associate researcher at China Medical University Hospital, shared his experience in using synthetic biology entrepreneurship; iGEM Asian Ambassador Chen Hong explained some new systems and shared experience on behalf of iGEM. We gained tons of new ideas from these lectures, which had great influences on our next work.

In the five-day conference, we also had a 20-minute English presentation and a poster exhibition. It was the first time to present our project frankly to iGEMers from Asia, and a good opportunity to practice for the final Jamboree. At the end of the exhibition, professor Yang Wen-Ming from the field of molecular biology at National Chung Hsing University gave a comment on our presentation. Through the conference, we also made suggestions to other Asian teams, longing for more cooperation opportunities.

This conference has provided us tremendous new ideas to improve our project and final presentation.

SUSTech iGEM

This year, the SUSTech iGEM team used the microfluidic chip as the core carrier to build a complete hardware platform, which was very similar to the idea of XMU-China team in hardware design. On this basis, these two teams had an online communication on issues related to microfluidic chips.

SUSTech used high-precision and single-channel chip tubes to achieve cell sorting functions, with a peristaltic pump that provides driving force, and a flow meter that filters and encapsulates cell droplets. These devices greatly improve the precision of the entire device.

XMU-China also made a relevant introduction about the design of microfluidic chips. What's more, our team additionally answered questions about how to control liquid flow in the centrifugal force-driven microfluidic chip.

Figure 1:The design of microfluidic chips of SUSTech iGEM(left) and XMU-China(right).

In terms of the project, we believe SUSTech iGEM team's using chips to study the interaction between cells is very innovative. From our professional perspective, we gave some advice on cell signaling pathways and cell-cell interactions, and recommended several related articles to them, hoping to help them. Based on our project, they raised some questions about whether our cell-free system belongs to the field of synthetic biology. We put forward our views, and the two parties had a futher discussion. We believe that synthetic biology is not a simple use of biobrick to construct gene loops. Instead, it should be a broader definition. We think that standard components are like building blocks, but it is up to people to decide what items should be joint together, rather than building blocks themselves. Therefore, biobrick can be used to build systems outside the genetic loop.

In addition, both of us exchanged ideas on the Art&Design. We carried out visual design communications with SUStech on aspects involving poster, Wiki and PPT. We showed them our methods and ideas in visual designing, including drawing graphics of experimental process and other details.

At CCiC, we are very delighted to find that BIT projects have a lot of intersections with us. We have similar representations.Therefore, our team communicated with them in the following three aspects.

The first is the exchange of ideas on the subject. We have had a two-hour telephone conversation with them. On the phone, we exchanged the purpose of the project and some detailed designs, introduced our own project to them and proposed some questions for each other.

Secondly, we asked the BIT team about a difficulty in the experimental program. I am very happy that they are effective. We proved that our aptamers and complementary chains are combined. We gave the results back to them and gave them suggestions of improvement. In addition, we also exchanged some experimental methods and protocol.

Finally, in order to prove the feasibility of their respective experimental designs, we mailed some materials to each other and successfully verified the effectiveness of the other's combination, which is also a very effective part of our cooperation.

The experimental results are shown in the figure below. Well 1: Aptamers, Well 2: Complementary chains, Well 3: Aptamers + Complementary chains. As can be seen from the figure, the electrophoresis speed of the combined DNA is slower than that of the aptamer and the complementary strand, and it is indeed combined.

On July 31, 2018, at the 6th Asia-Pacific iGEM Conference hosted by National Chung Hsing University, Taichung,we met with NAU-CHINA team and established a deep friendship. Since then, our two teams have exchanged and discussed "how to build a team."

The professor affirmed the feasibility of our project and gave us some invaluable advice. Under the suggestion of Professor Zhu Zhi, we chose EpCAM and aptamer SYL3C. In addition, we are very grateful to Professor Zhu for her support on many instruments and materials provided during our experiments.

First we discussed "what kind of team should we build?" The International Genetic Engineering Machine Competition is a creative synthetic biology event and an interdisciplinary competition involving cross-disciplinary fields in mathematics, computers, statistics and so on. So our team should be a creative, comprehensive synthetic biology team. Our team members should be creative and comprehensive students. Most students of our two teams have multiple roles in the team.

Second, we discussed "how to find new partners." After we know what kind of team we want to build, we could find the companions we need. First, we have presentations, exhibitions, lab activities, etc. to make students realize that, what kind of the team they will join in and what is the team building for? what role they will be in the team, and what should be done by themselves? In order to join this team and make this team better, how should them prepare and learn? Then we divide the interviewers into different groups according to different division of labor and their wishes. Different groups determine the way of assessment according to the division of labor. For example, the number module is tested by some models, and the art team performs preliminary selection through some art production tasks... We will give priority to students with multiple skills.

Third, we communicated on how to train the new team members in the early stages of team formation. First of all, we will introduce our team and International Genetic Engineering Machine Competition through presentations, posters, WeChat pushes, etc., and also convey our requests for new recruits. We will then train the interviewees in groups and we will select them during the training process. Our training is mainly to share the learning materials for students and let them learn by themselves.

Our two teams made recommendations on each other for training new members. They suggested us add a literature reading part to the training to teach students how to read literatures and how to extract important information from the literature. Then they shared with us some materials for teaching students to read the literature. Their advice has helped us a lot, and we plan to adopt their suggestions to improve this year's training. At the same time, we also suggest that they increase the opportunities for students to practice, let the students design their own topics, increase their free space, make them more creative and have more new ideas. In addition, we exchanged our training materials on modeling, hardware, software, art and web design with each other.

Finally, we communicated with the daily management of the team. At the beginning of each team, all the students will participate in the design of the project and conduct laboratory training. After the preliminary determination of the subject, the students were divided into different groups, responsible for different labors like experiments, modeling, human practice and so on. We hold group meetings every week, each group reports the progress of the project, and different groups communicate with each other. In addition, we will also hold regular entertainment activities such as picnics, parties, etc. to relax and enhance team friendship. Both of our teams have one thing in common. Most members are in multiple roles, which better strengthens the communication between groups. It shows that iGEM competition is a common combination. There are different groups in this team. Everyone is not working separately, but is always intimately connected.

Through the exchanges with the NAU-iGEM team, Our team has a better construction model, and we have more experience in the follow-up training this year. Thanks to the NAU team for their help.

This year XMU-china team and H14Z1_hangzhou team's cooperation can be divided into the following two parts:

First, XMU-China team helped and instructed H14Z1_hangzhou team to fill out the judging form, and to offer some advice about the relevant representations of one of a previous year biobricks to meet the bronze standard four. In addition, we also assisted H14Z1_hangzhou to complete the design of the first level menu and the second level menu in the navigation bar, so that their results can be better displayed in front of the judges.

Second, XMU-China and H14Z1_hangzhou assumed the task of the experimental representation each other. H14Z1_hangzhou helped XMU-China measure the fluorescence characterization data of BBa_K2623025. XMU-China helped H14Z1_hangzhou complete the experimental curve of the number of engineering bacteria in the process of fermenting yoghurt with the change of time.

The experimental steps and results are as following:

Experimental steps:

1. The first day, Lactobacillus NZ9000 and GMCA respectively were inoculated into 50ml GM17 medium, fermented at 30 ℃ overnight (about 12h). (Place them in a 30 ℃ incubator, no shaker shocks required).

2. Next day, fresh milk was heated to about 55-60 degrees Celsius, and then adding some white sugar, 3000rpm stirring for 10 minutes mixing. Divided them into 250ml shaker, and the volume of liquid in every shaker is 100ml.

3. Sterilize the above solution at 90 ℃ for 5min, then cool it to 30 degrees Celsius to prepare for inoculation and fermentation. Then collect Lactobacillus acidophilus in shaker and clean them 1 times with sterile water. Add Lactobacillus acidophilus to the sample making the initial OD is 0.1 (600nm). Then let them ferment statically in the 250ml shaker at 30 ℃ (Place them in the 30 ℃ incubator, without shaker vibration)

4. Measure the OD value every two hours and count the corresponding number of cells to draw the curve. Simultaneously, test the pH of the solution (Measure for 14 hours)

Experimental results:

Table 1 The comparison of biomass and product formation in two different yogurt

Cell number ( /ml) GSH (mg/L) SAM (mg/L)
Strain 6h 12h 6h 12h 6h 12h
L. lactis NZ9000 3.5x 107 1.1 x 109 0.0 0.0 0.3 0.8
L. lactis NZ9000/pGMcA 3.1x 107 1.0 x 109 4.5 11.8 1.5 2.6

2018 is the first year for FJNU-China team to participate in the iGEM competition, yet the teams competing in the first year are often lack of the relevant experience and grasp of the subject scale.

Since XMU-China team and FJNU-China are the only two teams in Fujian province in China, it is incumbent upon XMU-China team to instruct FJNU-China team and communicate with them to make the team fell they are parts of iGEM family as soon as possible.

In August, we held the Conference of Fujian Province iGEMer Community in China,and at the meeting, the both teams presented their own topics. Based on the topic of ideas, gene circuit design, experimental programs and practical application level, XMU-China put forward their own ideas, which helped FJNU-China better prepare for the competition and explain the ideas of their subject. After the meeting, we introduced them to the hospital's dermatologist and led them to visit the Xiamen Environmental Protection Bureau.

At the end of the competition, XMU-China provided years of experience to help FJNU-China to complete the safety form and judging form.

In the brainstorming stage, in order to solve the problem of preservation of Cas12a protein in microfluidic chips in hardware design, our team retrieved a large number of topics related to protein preservation in previous years. Finally inspired by 2017 TU_Delft's TDPs project and improving the characterization method. But it costed a lot of our time and efforts. After communicating with the team SYSU_Software in September, we asked them to quickly and accurately search for the previous project. They upgraded the functions of their software to cater our needs. We compared the information obtained by the manual search and the information obtained through the software search, and found that the software can be well positioned to the part of the experimental plan we need, which is equivalent to the accuracy of the manual search, but the work efficiency is improved and the work efficiency is lowered. After obtaining the results, we made recommendations on the user-friendliness of the software's user interface operations and the search algorithms can be optimized later.

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This year, the Tianjin team in China worked on Clock Mechanisms, which would utilize some circadian clock proteins discovered in Synechococcus elongatus (strain PCC 7942) (Anacystis nidulans R2).

We XMU-China also made efforts in Circadian clock “Kai”. Unfortunately, three kinds of protein which we used can not be produced in our lab, because the DNA ordered from IDT was disabled.

However, when we talked about circadian clock with members in Tianjin Team, we were lucky enough to be informed that they could provide us with the DNA we needed. Thanks to their donations, our study on rebuilding circadian clock “Kai” in E.coli can proceed.

More details about Tianjin’s work can be found in https://2018.igem.org/Team:Tianjin

This year we have newsletters with other iGEM teams from schools of all over the world, like Boston University (BU), Technische Universiteit Delft(TUDelft),TUST, ECUST, JNU and so on. Those letters are precious and beneficial since they enable different teams to share projects and views with each other, especially on topics like team information, team introduction, project description, etc. There is a link that you can see the new version of the newsletters. Through this platform, more people could learn about our projects and we have more opportunities to communicate and cooperate with the external world.