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+ | </style> | ||
+ | <style type="text/css"> | ||
+ | .Jnav{position: fixed;top: 17px;opacity: 1;background-color: rgba(255, 255, 255, 0.2); width: 100%;z-index:999;} | ||
+ | .Jnav a{text-decoration: none!important;color:#039be5;} | ||
+ | .Jnavtitle{float: right;width: 10%;text-align: center;padding: 1em 0;} | ||
+ | .Jnavdrag{position: relative;width: 100%;} | ||
+ | .Jnavdrag>ul{position: absolute;top: 0;width: 100%;border-radius: 5px;background-color: white;transition: all .4s ease-in-out;opacity: 0;} | ||
+ | </style> | ||
+ | <script type="text/javascript"> | ||
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+ | </script> | ||
+ | <div class="Jnav"> | ||
+ | <div class="Jnavtitle"> | ||
+ | <a href="https://2018.igem.org/Team:Jiangnan/Safety">Safety</a> | ||
+ | </div> | ||
+ | <div class="Jnavtitle"> | ||
+ | <a href="https://2018.igem.org/Team:Jiangnan/Hardware">Hardware</a> | ||
+ | </div> | ||
+ | <div class="Jnavtitle" onmouseover="Jnavshow(this)" onmouseleave="Jnavhide(this)"> | ||
+ | <a href="https://2018.igem.org/Team:Jiangnan/Team">Team</a> | ||
+ | <div class="Jnavdrag"> | ||
+ | <ul> | ||
+ | <li><a href="https://2018.igem.org/Team:Jiangnan/Team">Team Members</a></li> | ||
+ | <li class="divider"></li> | ||
+ | <li><a href="https://2018.igem.org/Team:Jiangnan/Attributions">Attribution</a></li> | ||
+ | <li class="divider"></li> | ||
+ | <li><a href="https://2018.igem.org/Team:Jiangnan/Collaborations">Collaboration</a></li> | ||
+ | </ul> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="Jnavtitle" onmouseover="Jnavshow(this)" onmouseleave="Jnavhide(this)"> | ||
+ | <a href="https://2018.igem.org/Team:Jiangnan/Human_Practices">Human Practice</a> | ||
+ | <div class="Jnavdrag"> | ||
+ | <ul> | ||
+ | <li><a href="https://2018.igem.org/Team:Jiangnan/Human_Practices">Overview</a></li> | ||
+ | <li class="divider"></li> | ||
+ | <li><a href="https://2018.igem.org/Team:Jiangnan/Silver">Silver</a></li> | ||
+ | <li class="divider"></li> | ||
+ | <li><a href="https://2018.igem.org/Team:Jiangnan/Entrepreneurship">Gold</a></li> | ||
+ | <li class="divider"></li> | ||
+ | <li><a href="https://2018.igem.org/Team:Jiangnan/Public_Engagement">Pulic Engagement</a></li> | ||
+ | <li class="divider"></li> | ||
+ | <li><a href="https://2018.igem.org/Team:Jiangnan/Entrepreneurship">Entrepreneurship</a></li> | ||
+ | </ul> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="Jnavtitle"> | ||
+ | <a href="https://2018.igem.org/Team:Jiangnan/Model">Model</a> | ||
+ | </div> | ||
+ | <div class="Jnavtitle" onmouseover="Jnavshow(this)" onmouseleave="Jnavhide(this)"> | ||
+ | <a href="https://2018.igem.org/Team:Jiangnan/Notebook">Notebook</a> | ||
+ | <div class="Jnavdrag"> | ||
+ | <ul> | ||
+ | <li><a href="https://2018.igem.org/Team:Jiangnan/Notebook">Lab Book</a></li> | ||
+ | <li class="divider"></li> | ||
+ | <li><a href="https://2018.igem.org/Team:Jiangnan/Protocol">Protocol</a></li> | ||
+ | </ul> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="Jnavtitle" onmouseover="Jnavshow(this)" onmouseleave="Jnavhide(this)"> | ||
+ | <a href="https://2018.igem.org/Team:Jiangnan">Project</a> | ||
+ | <div class="Jnavdrag"> | ||
+ | <ul> | ||
+ | <li><a href="https://2018.igem.org/Team:Jiangnan/Background">Background</a></li> | ||
+ | <li class="divider"></li> | ||
+ | <li><a href="https://2018.igem.org/Team:Jiangnan/Design">Design</a></li> | ||
+ | <li class="divider"></li> | ||
+ | <li><a href="https://2018.igem.org/Team:Jiangnan/Demonstrate">Demonstration</a></li> | ||
+ | <li class="divider"></li> | ||
+ | <li><a href="https://2018.igem.org/Team:Jiangnan/Results">Result</a></li> | ||
+ | <li class="divider"></li> | ||
+ | <li><a href="https://2018.igem.org/Team:Jiangnan/Parts">Part</a></li> | ||
+ | </ul> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="navlogo" style="float: left;width: 20%;text-align: center;"> | ||
+ | <a href="https://2018.igem.org/Team:Jiangnan"><img src="https://static.igem.org/mediawiki/2018/d/d7/T--Jiangnan--igemJN_logo.png" style="width: 3em;"></a> | ||
+ | </div> | ||
</div> | </div> | ||
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− | <div | + | <div style="width:100%;background-color: #f0ebea"> |
− | < | + | <img src="https://static.igem.org/mediawiki/2018/5/59/T--Jiangnan--design_top.png" width="100%" style="margin-top: -7em;"> |
− | + | <div class="row"> | |
− | + | <div class="col s1" style="position: absolute;right: 0px; top: 30%;padding-right: 0;"> | |
− | < | + | <img src="https://static.igem.org/mediawiki/2018/7/71/T--Jiangnan--design_rightcircle1.png" width="100%"> |
− | < | + | </div> |
− | </ | + | |
</div> | </div> | ||
+ | <div class="row" style="position: relative; margin-top: 5em;margin-bottom: 5em;"> | ||
+ | <div class="col s10 offset-s1"> | ||
+ | <h4>Broad Spectrum</h4> | ||
+ | </div> | ||
+ | <div class="col s10 offset-s1"> | ||
+ | <p>The key for a cell line to be sensitive to the infection of a broad spectrum of viruses is to have the corresponding receptors expressed on the cell surface. We established an association between virus Baltimore subtyping and cell surface receptors mediating virus infection, and found that given the current sensitivity spectrum of our chassis cell line MDBK, only Nectin4 and TfR need to be artificially expressed on cell surface to make it sensitive to, in principle, all commonly encountered animal viruses. The sequence design is as below, where the sequences encoding each receptor are inserted into the designed cassette,respectively, and the proteins are expressed after cell transfection.</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="row" style="position: relative;margin-top: 5em;margin-bottom: 5em;"> | ||
+ | <div class="col s8 offset-s3"> | ||
+ | <img src="https://static.igem.org/mediawiki/2018/3/33/T--Jiangnan--design_procedure.png" width="100%"> | ||
+ | </div> | ||
+ | <div class="col s3" style="position: absolute;left: 0;bottom: -60%;padding-left: 0;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2018/9/92/T--Jiangnan--design_leftcircle1.png" width="100%"> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="row" style="position: relative;margin-top: 5em;margin-bottom: 5em;"> | ||
+ | <div class="col s8 offset-s3" style="text-align: right;"> | ||
+ | <h4>Suspension Culture</h4> | ||
+ | <p>Through comparing two sets (CHO and BHK-21) of transcriptome of suspended and adherent cells with the same origin, we obtained a list of candidate genes responsible for cell suspension. Following computational modeling of these genes, we identified the target gene “PABPC 1” and conducted wet lab experiments to verify our assumption. The gene regulatory network is constructed as below.</p> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="row" style="position: relative;margin-top: 5em;margin-bottom: 5em;"> | ||
+ | <div class="col s12" style="text-align: center;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2018/5/58/T--Jiangnan--design_hightiter.png" width="40%"> | ||
+ | </div> | ||
+ | <div class="col s1" style="position: absolute;right: 0;padding-right: 0;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2018/6/6e/T--Jiangnan--design_rightcircle2.png" width="100%"> | ||
+ | </div> | ||
+ | </div> | ||
+ | <div class="row" style="position: relative;margin-top: 5em;margin-bottom: 5em;"> | ||
+ | <div class="col s10 offset-s1"> | ||
+ | <h4>High Titer</h4> | ||
+ | <p>We conducted two sets of work to increase virus titer.</p> | ||
+ | <p>First, we constructed the computational network and identified the key gene (IRF7) responsible for virus multiplication, which is shown as below.</p> | ||
+ | </div> | ||
+ | <div class="col s11" style="text-align: center;"> | ||
+ | <img src="https://static.igem.org/mediawiki/2018/d/da/T--Jiangnan--design_suspension.png" width="40%"> | ||
+ | </div> | ||
+ | <div class="col s10 offset-s1"> | ||
+ | <p>Second, we established an approach, namely cold atmospheric plasma, with the device manufactured to further increase virus titre. Plasma is the fourth fundamental state of matter, and a cocktail of ions, free electrons, reactive species, etc. Plasma may increase virus titre by increasing the permeability of cell membrane or stimulating cell receptors mediating virus internalization.</p> | ||
+ | </div> | ||
</div> | </div> | ||
+ | </div> | ||
</html> | </html> |
Revision as of 10:26, 17 October 2018
![](https://static.igem.org/mediawiki/2018/5/59/T--Jiangnan--design_top.png)
![](https://static.igem.org/mediawiki/2018/7/71/T--Jiangnan--design_rightcircle1.png)
Broad Spectrum
The key for a cell line to be sensitive to the infection of a broad spectrum of viruses is to have the corresponding receptors expressed on the cell surface. We established an association between virus Baltimore subtyping and cell surface receptors mediating virus infection, and found that given the current sensitivity spectrum of our chassis cell line MDBK, only Nectin4 and TfR need to be artificially expressed on cell surface to make it sensitive to, in principle, all commonly encountered animal viruses. The sequence design is as below, where the sequences encoding each receptor are inserted into the designed cassette,respectively, and the proteins are expressed after cell transfection.
![](https://static.igem.org/mediawiki/2018/3/33/T--Jiangnan--design_procedure.png)
![](https://static.igem.org/mediawiki/2018/9/92/T--Jiangnan--design_leftcircle1.png)
Suspension Culture
Through comparing two sets (CHO and BHK-21) of transcriptome of suspended and adherent cells with the same origin, we obtained a list of candidate genes responsible for cell suspension. Following computational modeling of these genes, we identified the target gene “PABPC 1” and conducted wet lab experiments to verify our assumption. The gene regulatory network is constructed as below.
![](https://static.igem.org/mediawiki/2018/5/58/T--Jiangnan--design_hightiter.png)
![](https://static.igem.org/mediawiki/2018/6/6e/T--Jiangnan--design_rightcircle2.png)
High Titer
We conducted two sets of work to increase virus titer.
First, we constructed the computational network and identified the key gene (IRF7) responsible for virus multiplication, which is shown as below.
![](https://static.igem.org/mediawiki/2018/d/da/T--Jiangnan--design_suspension.png)
Second, we established an approach, namely cold atmospheric plasma, with the device manufactured to further increase virus titre. Plasma is the fourth fundamental state of matter, and a cocktail of ions, free electrons, reactive species, etc. Plasma may increase virus titre by increasing the permeability of cell membrane or stimulating cell receptors mediating virus internalization.