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− | <p> | + | <p>Since collaboration is one of the hallmarks and core concepts of iGEM, cooperating with Chinese iGEMers has always been our tradition and focus since we set up team NJU-China. Through collaborations and partnerships with other Chinses iGEM team, we share our idea, provide assistance within our capability, promote our friendships, and strengthen our understanding of synthetic biology. And on this page, we would like express our heartfelt gratitude towards all the teams below that we have kept sound cooperative relationship with this year. Furthermore, we expect that all of these will pave the way for a deeper cooperation and are looking forward to build friendships with more iGEMers around the world in the near future.</p> |
− | + | ||
</div> | </div> | ||
<div class="divider"></div> | <div class="divider"></div> | ||
+ | <ul class="collapsible" data-collapsible="expandable"> | ||
+ | <li> | ||
+ | <div class="collapsible-header active"><i class="material-icons">filter_drama</i>Collaboration with SYSU-Software</div> | ||
+ | <div class="collapsible-body"> | ||
+ | <p>In order to design short hairpin RNA (shRNA) conform to demands in our experiments, we cooperated with SYSU-Software from Sun Yat-sen University to develop a corresponding software. We provided algorithms and design requirements, and then SYSU-Software wrote this delicate software for us. With their help, we successfully designed K-ras shRNA, whose sequence and predicted structure are shown below (Fig. 1). We validated that this shRNA was able to down-regulate the expression level of K-ras in A549 effectively. We thank SYSU-Software for receiving us with attention and for such strong contribution to our work.</p> | ||
+ | <img src="" class="responsive-img" align="right"> | ||
+ | <img src="" class="responsive-img"> | ||
+ | <div align="center">Figure. 1 Sequence and predicted structure of K-ras shRNA</div> | ||
+ | </div> | ||
+ | </li> | ||
+ | <li> | ||
+ | <div class="collapsible-header active"><i class="material-icons">filter_drama</i>Collaboration with NUDT_CHINA</div> | ||
+ | <div class="collapsible-body"> | ||
+ | <p>Since team NUDT_CHINA met some troubles in assessing the efficiency of the single guide RNAs designed for their experiments, we performed a Cas9-digest assay to assist them validating the effectiveness of their sgRNAs. Also, we helped them to optimize their own protocol for in vitro expression of sgRNAs, so that their project could be performed more smoothly.</p> | ||
+ | <p>With the expressing plasmids provided by team NUDT_CHINA, in vitro transcription based on T7 RNA polymerase was conducted to obtain enough amount of sgRNAs. Two standard sgRNAs, with the binding efficiency of 20% and 70% were also transcribed to evaluate the efficiency of sgRNAs. Once transcribed, purified Cas9 protein was used to digest target DNA (provided by NUDT_CHINA team) with the guidance of sgRNAs.</p> | ||
+ | <p>Results showed that when transcribed correctly (Fig. 2A), sgR-1 and sgR-4 could successfully mediate the digestion of target DNA by Cas9 protein, while the negative control adding no sgRNA showing no digestion output. The binding efficiency was estimated to be between 20% and 70%, while the binding efficiency of sgR-1 is significantly higher than sgR-4 (Fig. 2B). Those results indicated that sgR-1 could be a better choice to be used in their project.</p> | ||
+ | <img src="" class="responsive-img" align="right"> | ||
+ | <img src="" class="responsive-img"> | ||
+ | <div align="center">Figure. 1 Sequence and predicted structure of K-ras shRNA</div> | ||
+ | <p>In addition, we'd like to express our sincere thanks to NUDT_CHINA for their generous help in Nanosight, enabling us to have a much more precise determination of the quantity and size of exosomes we collected. You can click on this link to access to the detailed information: (result中Nanosight部分的链接)</p> | ||
+ | </div> | ||
+ | </li> | ||
+ | <li> | ||
+ | <div class="collapsible-header active"><i class="material-icons">filter_drama</i>Collaboration with SCUT-CHINA</div> | ||
+ | <div class="collapsible-body"> | ||
+ | <p>We much appreciate that SCUT-China from South China University of Technology provided vigorous support and supplement in our modeling. For more information: (Wiki-Model的链接)</p> | ||
+ | <img src="" class="responsive-img" align="right"> | ||
+ | </div> | ||
+ | </li> | ||
+ | <li> | ||
+ | <div class="collapsible-header active"><i class="material-icons">filter_drama</i>Collaboration with JSNU-China</div> | ||
+ | <div class="collapsible-body"> | ||
+ | <p>Since JSNU-China from Jiangsu Normal University confronted some troubles in their research on neuroblastoma cells and we had done some serious experiments in this field, we offered them cells and gave them some pieces of advice.</p> | ||
+ | <img src="" class="responsive-img" align="right"> | ||
+ | </div> | ||
+ | </li> | ||
+ | </ul> | ||
</div> | </div> | ||
</main> | </main> |
Revision as of 13:27, 19 October 2016
Since collaboration is one of the hallmarks and core concepts of iGEM, cooperating with Chinese iGEMers has always been our tradition and focus since we set up team NJU-China. Through collaborations and partnerships with other Chinses iGEM team, we share our idea, provide assistance within our capability, promote our friendships, and strengthen our understanding of synthetic biology. And on this page, we would like express our heartfelt gratitude towards all the teams below that we have kept sound cooperative relationship with this year. Furthermore, we expect that all of these will pave the way for a deeper cooperation and are looking forward to build friendships with more iGEMers around the world in the near future.
-
filter_dramaCollaboration with SYSU-Software
In order to design short hairpin RNA (shRNA) conform to demands in our experiments, we cooperated with SYSU-Software from Sun Yat-sen University to develop a corresponding software. We provided algorithms and design requirements, and then SYSU-Software wrote this delicate software for us. With their help, we successfully designed K-ras shRNA, whose sequence and predicted structure are shown below (Fig. 1). We validated that this shRNA was able to down-regulate the expression level of K-ras in A549 effectively. We thank SYSU-Software for receiving us with attention and for such strong contribution to our work.
Figure. 1 Sequence and predicted structure of K-ras shRNA -
filter_dramaCollaboration with NUDT_CHINA
Since team NUDT_CHINA met some troubles in assessing the efficiency of the single guide RNAs designed for their experiments, we performed a Cas9-digest assay to assist them validating the effectiveness of their sgRNAs. Also, we helped them to optimize their own protocol for in vitro expression of sgRNAs, so that their project could be performed more smoothly.
With the expressing plasmids provided by team NUDT_CHINA, in vitro transcription based on T7 RNA polymerase was conducted to obtain enough amount of sgRNAs. Two standard sgRNAs, with the binding efficiency of 20% and 70% were also transcribed to evaluate the efficiency of sgRNAs. Once transcribed, purified Cas9 protein was used to digest target DNA (provided by NUDT_CHINA team) with the guidance of sgRNAs.
Results showed that when transcribed correctly (Fig. 2A), sgR-1 and sgR-4 could successfully mediate the digestion of target DNA by Cas9 protein, while the negative control adding no sgRNA showing no digestion output. The binding efficiency was estimated to be between 20% and 70%, while the binding efficiency of sgR-1 is significantly higher than sgR-4 (Fig. 2B). Those results indicated that sgR-1 could be a better choice to be used in their project.
Figure. 1 Sequence and predicted structure of K-ras shRNAIn addition, we'd like to express our sincere thanks to NUDT_CHINA for their generous help in Nanosight, enabling us to have a much more precise determination of the quantity and size of exosomes we collected. You can click on this link to access to the detailed information: (result中Nanosight部分的链接)
-
filter_dramaCollaboration with SCUT-CHINA
We much appreciate that SCUT-China from South China University of Technology provided vigorous support and supplement in our modeling. For more information: (Wiki-Model的链接)
-
filter_dramaCollaboration with JSNU-China
Since JSNU-China from Jiangsu Normal University confronted some troubles in their research on neuroblastoma cells and we had done some serious experiments in this field, we offered them cells and gave them some pieces of advice.