Team:NJU China/Data Page

From 2013.igem.org

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</head>
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<body>
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<div class="container">
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<!--Start NavBar-->
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<ul id="nav">
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    <li><a href="https://2013.igem.org/Team:NJU_China">Home</a></li>
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    <li><a href="https://2013.igem.org/Team:NJU_China/Project">Project</a>
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        <ul>
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  <li><a href="http://https://2013.igem.org/Team:NJU_China/Project#Overview">Overview</a></li>
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          <li><a https://2013.igem.org/Team:NJU_China/Project#Chassis">Chassis</a></li>
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          <li><a https://2013.igem.org/Team:NJU_China/Project#Targeting Module">Targeting Module</a></li>
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          <li><a https://2013.igem.org/Team:NJU_China/Project#Killing Module">Killing Module</a></li>
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          <li><a https://2013.igem.org/Team:NJU_China/Project#Achievement">Achievement</a></li>
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        </ul>
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    </li>
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    <li><a href="https://2013.igem.org/Team:NJU_China/Modeling">Modeling</a>
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    </li>
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    <li><a href="https://2013.igem.org/Team:NJU_China/Human Practice">Human Practice</a>
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        <ul>
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  <li><a href="http://https://2013.igem.org/Team:NJU_China/Project#Free debate">Free debate</a></li>
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          <li><a https://2013.igem.org/Team:NJU_China/Project#Reach the unreached">Reach the unreached</a></li>
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          <li><a https://2013.igem.org/Team:NJU_China/Project#Workshop">Workshop</a></li>
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          <li><a https://2013.igem.org/Team:NJU_China/Project#Collaboration">Collaboration</a></li>
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        </ul>
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    </li>
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    <li><a href="https://2013.igem.org/Team:NJU_China/Wet lab">Wet lab</a>
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        <ul>
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  <li><a href="http://https://2013.igem.org/Team:NJU_China/Project#Protocol">Protocol</a></li>
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          <li><a https://2013.igem.org/Team:NJU_China/Project#Reach the unreached">Lab notes</a></li>
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          <li><a https://2013.igem.org/Team:NJU_China/Project#Workshop">Data Page</a></li>
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          <li><a https://2013.igem.org/Team:NJU_China/Project#Collaboration">Parts</a></li>
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        </ul>
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    </li>
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    <li><a href="https://2013.igem.org/Team:NJU_China/Safety">Safety</a>
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        <ul>
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  <li><a href="http://https://2013.igem.org/Team:NJU_China/Project#Protocol">Safety form</a></li>
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          <li><a https://2013.igem.org/Team:NJU_China/Project#Reach the unreached">Virus-related safety</a></li>
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        </ul>
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    </li>
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    <li><a href="https://2013.igem.org/Team:NJU_China/Team">Team</a></li>
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    <li><a href="https://2013.igem.org/Team:NJU_China/Extras">Extras</a>
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            <ul>
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  <li><a href="http://https://2013.igem.org/Team:NJU_China/Project#Protocol">Judging criteria</a></li>
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          <li><a https://2013.igem.org/Team:NJU_China/Project#Reach the unreached">Attribution</a></li>
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          <li><a https://2013.igem.org/Team:NJU_China/Project#Reach the unreached">Acknowledgement</a></li>
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        </ul>
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    </li>
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</ul>
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<!--End NavBar-->
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<header>
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<h1>Liver: <span>For liver targeting, we need to first find a protein specifically recognize hepatic cells. Since Heptitis B virus can infect hepatic cells distinctively, and from recent study[1], we knew that HBV recognizes the hepatic cells via the interaction between the pre-S1 of the HBV envelop protein and NTCP receptor of the hepatic cells. We tried to engineer the pre-S1 from HBV envelope protein to the lamp 2b.
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Therefore we cloned the pre-S1 into lamp 2b, and we choose pcDNA 3.1(+) as our backbone.</span>
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<img style="float:left" src="https://static.igem.org/mediawiki/2013/d/d9/Pcdna3.1_pres1_small.png">
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</br><img width="460px"; height="300px" src="https://static.igem.org/mediawiki/2013/3/3e/%E8%AF%86%E5%88%ABpre_s1_small.png">
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</h1>
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</header>
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</br>
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<section>
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<p>Results for liver targeting:</BR>
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To produce the exosomes that have pre-S1 on their surface for liver targeting, we first transfected the exosome-producing cells, HEK 293T cells, with the plasmid encoding the fusion protein of lamp 2b and pre-S1 peptide.</p>
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<p>1. Exosome morphology under SEM and TEM</br>
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The exosomes produced by the transfected HEK 293T cells was collected 24h post transfection. The morphology and diameter of the exosomes were examined by both SEM and TEM.</br>
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As shown in Fig.1 and Fig.2, the diameter of the exosomes is around 50nm and it was round.</p>
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<p>2. In vitro evidence for the entry of pre-S1 exosomes into the hep G2 cell</br>
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As shown in Fig.3, by labeling the exosomes with DiI-C16 (red) and hep G2 nucleus with DAPI(blue), it can be seen that the exosomes successfully get into the hep G2 cells.</p>
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<img width="500px"; height="500px" src="https://static.igem.org/mediawiki/2013/c/c3/NjuLiver.png">
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<p>Fig.3 Confocal microscopy image of the internalization of fluorescently labeled MVs into hep  cells.  293T cells were labeled with DiI-C16 (red) and then cultured in RPMI 1640 medium supplemented with 10% FBS. After 4 hr, the supernatants were collected and centrifuged to harvest exosomes.  293T exosomes were resuspended in MCDB-131 medium and incubated with hep G2 cells at 37􏰀C. After incubation for 2 hr, hep G2 cells were washed, fixed, and observed under confocal microscopy.</p>
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<p>3. In vivo evidence for the entry of pre-S1 exosomes into the liver</p>
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</section>
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</div><!-- /container -->
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</body>
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</html>
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Revision as of 15:57, 27 September 2013

<frameset rows="20%,*" border="0px"> <frame> Hello World!</frame> <frameset cols="20%,*"> <frame name="frame1" src="left.html"/> <frame name="frame2" src="all.html"/> </frameset> </frameset>

Retrieved from "http://2013.igem.org/Team:NJU_China/Data_Page"