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<div id="float01" class="cur"> | <div id="float01" class="cur"> | ||
<div class="h1">Attributions</div> | <div class="h1">Attributions</div> | ||
− | <p>Our project | + | <p>Our project was designed by <b>Zhujun Xia</b> and all circuits were designed by <b>Zhujun Xia</b>, <b>YinQing |
Zeng</b> and | Zeng</b> and | ||
<b>Mo Qiqin</b>.</p> | <b>Mo Qiqin</b>.</p> | ||
<div class="h2">Wet Lab</div> | <div class="h2">Wet Lab</div> | ||
<div class="h3">Gene Editing</div> | <div class="h3">Gene Editing</div> | ||
− | <p> | + | <p><i>sifA</i> was knocked out by <b>Zhujun Xia</b>, <b>Zhuoqi Huang</b> and <b>Wenxin Hu</b> |
also gave | also gave | ||
assistance.</p> | assistance.</p> | ||
<div class="h3">Plasmid Construction</div> | <div class="h3">Plasmid Construction</div> | ||
− | <p>Intracellular Environment-Dependent Circuits:</p> | + | <p><b>Intracellular Environment-Dependent Circuits:</b></p> |
− | <p>The intracellular environment-dependent circuits | + | <p>The intracellular environment-dependent circuits were designed by <b>Mo Qiqin</b> and <b>Zhujun Xia</b>.</p> |
− | <p>The intracellular environment-dependent circuits | + | <p>The intracellular environment-dependent circuits were constructed by <b>Mo Qiqin</b>. <b>Ruonan Tian</b> |
also gave | also gave | ||
assistance.</p> | assistance.</p> | ||
<br> | <br> | ||
− | <p>Chemical Control System:</p> | + | <p><b>Chemical Control System:</b></p> |
− | <p>The ATc | + | <p>The ATc induction chemical control system was constructed by <b>He Yang</b> and <b>Zhuoqi Huang</b> also |
gave | gave | ||
assistance.</p> | assistance.</p> | ||
− | <p>The TetR-eGFP fusion protein expression system for TetR expression | + | <p>The TetR-eGFP fusion protein expression system for TetR expression measurement was constructed by <b>He |
− | Yang</b> | + | Yang</b> and <b>Zhuoqi Huang</b> also gave assistance.</p> |
<br> | <br> | ||
− | <p>Targeting Specificity and Surface-Displaying:</p> | + | <p><b>Targeting Specificity and Surface-Displaying:</b></p> |
− | <p>RGD-inserted OmpA circuit clone and construction of Salmonella SL1344_RS05255 (OmpA coding | + | <p>RGD-inserted OmpA circuit clone and construction of <i>Salmonella</i> SL1344_RS05255 (OmpA coding |
− | sequence) | + | sequence) were completed by <b>YinQing Zeng</b> and <b>Lingyu Zhong</b>.</p> |
− | <p>The circuit of surface-display system | + | <p>The circuit of surface-display system was constructed by <b>Lingyu Zhong</b>. |
− | Optimation of OmpA in E. Coli | + | Optimation of OmpA in <i>E. Coli</i> was completed by <b>Zhujun Xia</b>.</p> |
<br> | <br> | ||
− | <p>Plasmid Standardization:</p> | + | <p><b>Plasmid Standardization:</b></p> |
− | <p>Plasmid standardization | + | <p>Plasmid standardization was completed by <b>Zhiqing Guo</b>, <b>Hangxi Fu</b> and <b>Zhuoqi Huang</b> by using 3A |
Assembly. | Assembly. | ||
</p> | </p> | ||
<br> | <br> | ||
− | <p>Plasmid Submission:</p> | + | <p><b>Plasmid Submission:</b></p> |
− | <p>Submitted plasmids (Pcs2-eGFP-GSDMD FL, Pcs2-eGFP-GSDMD-N275 and | + | <p>Submitted plasmids (Pcs2-eGFP-GSDMD FL, Pcs2-eGFP-GSDMD-N275 and two GSDMD full length mutations) |
− | + | were constructed by <b>Zhiqing Guo</b> and <b>Wenxin Hu</b>.</p> | |
− | <p>The description of submitted plasmids by ATP assay and cell phenotype via transfection | + | <p>The description of submitted plasmids by ATP assay and cell phenotype via transfection was designed |
and completed by <b>Zhujun Xia</b>. | and completed by <b>Zhujun Xia</b>. | ||
</p> | </p> | ||
<div class="h3">Bacterial Phenotype Verification</div> | <div class="h3">Bacterial Phenotype Verification</div> | ||
− | <p>Bacterial phenotype | + | <p>Bacterial phenotype verifications were designed and completed by <b>He Yang</b>, including the OD |
measurement | measurement | ||
and CFU measurement of chemical control system.</p> | and CFU measurement of chemical control system.</p> | ||
<div class="h3">Cell Phenotype Verification</div> | <div class="h3">Cell Phenotype Verification</div> | ||
− | <p>Cell phenotype verifications | + | <p>Cell phenotype verifications were designed and completed by <b>Zhujun Xia</b>, including the |
− | + | verification | |
− | of chemical control system, and targeting specificity, all | + | of chemical control system, and targeting specificity, all the related circuits and the demonstration of the final work system.</p> |
− | <div class="h3"> | + | <div class="h3">Biosafety Verification</div> |
− | <p>Safety verification | + | <p>Safety verification was designed and completed by <b>Zhujun Xia</b>, which demonstrated our project |
is safe | is safe | ||
enough to be used.</p> | enough to be used.</p> | ||
<div class="h3">InterLab</div> | <div class="h3">InterLab</div> | ||
− | <p>InterLab | + | <p>InterLab was conducted by <b>Xichen Rao</b>, <b>Zhujun Xia</b>, <b>Ruonan Tian</b> and <b>Heng Heng</b> |
− | also gave assistance | + | also gave assistance. |
− | + | Our data passed the check of the Measurement Committee successfully.</p> | |
<div class="h2">Dry Lab</div> | <div class="h2">Dry Lab</div> | ||
− | <p>Mathematical model and app | + | <p>Mathematical model and app about salmonella infection and GSDMD release were built by <b>Songtao |
− | Chen</b>. | + | Chen</b>.</p> |
− | ATc model which predicted the relationship between concentration of ATc and concentration of GSDMD | + | <p>ATc model which predicted the relationship between concentration of ATc and concentration of GSDMD |
− | + | was built by <b>Xichen Rao</b>.</p> | |
− | The modeling | + | <p>The modeling on adjusting the strength of RBS and ATc with fixed GSDMD-N concentration was completed |
− | by <b>Yini Miao</b>. | + | by <b>Yini Miao</b>. </p> |
− | The modeling of the promoter sifA and promoter | + | <p>The modeling of the promoter sifA and promoter ent was completed by <b>Ruonan Tian</b>. |
</p> | </p> | ||
<div class="h2">Software</div> | <div class="h2">Software</div> | ||
− | <p>The applet of | + | <p>The applet of "Bacterial Colony Counter" was designed and programmed by <b>Shuguang Wang</b>.</p> |
<div class="h2">Human Practice</div> | <div class="h2">Human Practice</div> | ||
− | <p>Collaboration and communication with other teams | + | <p>Collaboration and communication with other teams were completed by <b>Yinqing Zeng</b> and all the other team members.</p> |
− | Handicraft manufacture | + | <p>Handicraft manufacture was designed and completed by <b>Yinqing Zeng</b>.</p> |
− | Questionnaire preparation, the advertisement of synthetic biology and popular science articles | + | <p>Questionnaire preparation, the advertisement of synthetic biology and preparation of popular science articles |
− | + | were completed by <b>Heng Heng</b>.</p> | |
− | Two posters for Eurasian exchange meetings and CCIC | + | <p>Two posters for Eurasian exchange meetings and CCIC were completed by <b>Ruonan Tian</b>. |
</p> | </p> | ||
<div class="h2">Website</div> | <div class="h2">Website</div> | ||
− | <p>Our | + | <p>Our wiki framework was designed and programmed by <b>Shuguang Wang</b> and our artist were |
− | + | <b>Yuying Xiang</b> and <b>Kening Chen</b>.</p> | |
− | <p>Our <a href="https://wang-shuguang.github.io/">blog</a> | + | <p>Our <a href="https://wang-shuguang.github.io/">blog</a> was built by <b>Shuguang Wang</b>.</p> |
<div class="h2">Others</div> | <div class="h2">Others</div> | ||
− | <p>The plate reader machine operation | + | <p>The plate reader machine operation was conducted by <b>Mo Qiqin</b>.</p> |
− | + | <p>Chief cleaner of the laboratory was <b>Xichen Rao</b>. | |
</p> | </p> | ||
</div> | </div> | ||
<div id="float02"> | <div id="float02"> | ||
− | <div class="h1"> | + | <div class="h1">Acknowledgements</div> |
<p>For each parts of experiments, our appreciations are as follow:</p> | <p>For each parts of experiments, our appreciations are as follow:</p> | ||
<div class="h2">Cell lines</div> | <div class="h2">Cell lines</div> | ||
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Thanks to Prof. <b>Shan Li</b>’s laboratory, Huazhong Agricultural University, for sharing the cell | Thanks to Prof. <b>Shan Li</b>’s laboratory, Huazhong Agricultural University, for sharing the cell | ||
lines | lines | ||
− | of HEK293T, HELA, MCF7, iBMDM, our experiment can process successfully. | + | of HEK293T, HELA, MCF7, iBMDM, so that our experiment can process successfully. |
</p> | </p> | ||
− | <p> Thanks to Dr. <b>Feng Shao</b>’s laboratory, for sharing the cell line of GSDMD-/- HELA cells, | + | <p> Thanks to Dr. <b>Feng Shao</b>’s laboratory, for sharing the cell line of GSDMD<i>-/-</i> HELA cells, which |
provides | provides | ||
us great support. | us great support. | ||
Line 823: | Line 820: | ||
<p>Thanks to Prof. <b>Shan Li</b>’s laboratory, Huazhong Agricultural University, for sharing the | <p>Thanks to Prof. <b>Shan Li</b>’s laboratory, Huazhong Agricultural University, for sharing the | ||
strain of | strain of | ||
− | Salmonella enterica var. Typhimurium SL1344. | + | <i>Salmonella enterica</i> var. Typhimurium SL1344. |
</p> | </p> | ||
− | <p> Thanks to Dr. Chenli Liu from SIAT | + | <p> Thanks to Dr. <b>Chenli Liu</b> from SIAT CSynBER for generously sharing the material (Strain: <i>E.coli</i> K-12 MG1655 with |
− | mCherry in the genome) | + | mCherry in the genome). |
</p> | </p> | ||
<div class="h2">Gene Knock In and Knock Out</div> | <div class="h2">Gene Knock In and Knock Out</div> | ||
<p>Thanks to Prof. <b>Shan Li</b>’s laboratory, Huazhong Agricultural University, for providing the | <p>Thanks to Prof. <b>Shan Li</b>’s laboratory, Huazhong Agricultural University, for providing the | ||
protocol | protocol | ||
− | and material of gene editing based on two-step allelic exchange, | + | and material of gene editing based on two-step allelic exchange, which provides us with great help. |
− | + | ||
− | + | ||
− | + | ||
</p> | </p> | ||
+ | |||
<div class="h2">Plasmid Backbone</div> | <div class="h2">Plasmid Backbone</div> | ||
<p>Thanks to Prof. <b>Shan Li</b>’s laboratory, Huazhong Agricultural University, for sharing the | <p>Thanks to Prof. <b>Shan Li</b>’s laboratory, Huazhong Agricultural University, for sharing the | ||
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backbone (Pcs2-eGFP) to us.</p> | backbone (Pcs2-eGFP) to us.</p> | ||
<div class="h2">Sponsors</div> | <div class="h2">Sponsors</div> | ||
− | <p>Thanks to <b>GeneScript</b> and <b>IDT</b> for DNA sequence synthesis service</p> | + | <p>Thanks to <b>GeneScript</b> and <b>IDT</b> for DNA sequence synthesis service.</p> |
− | <p>Thanks to <b>SnapGene</b> and <b>MathWorks</b> for free software supporting</p> | + | <p>Thanks to <b>SnapGene</b> and <b>MathWorks</b> for free software supporting.</p> |
<div class="row"> | <div class="row"> | ||
<div class="col-xs-6 col-sm-6 col-md-6 col-lg-6"> | <div class="col-xs-6 col-sm-6 col-md-6 col-lg-6"> | ||
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<br> | <br> | ||
<br> | <br> | ||
− | <p>Thanks to College of Life Science and | + | <p>Thanks to <b>College of Life Science and Technology and Huazhong Agricultural University</b> for providing |
− | the site and fund for us. | + | the working site and fund for us. |
</p> | </p> | ||
<div style="width: 50%; margin: 10px auto"> | <div style="width: 50%; margin: 10px auto"> | ||
<img src="https://static.igem.org/mediawiki/2018/1/1e/T--HZAU-China--huanong.png" width=100% alt=""> | <img src="https://static.igem.org/mediawiki/2018/1/1e/T--HZAU-China--huanong.png" width=100% alt=""> | ||
</div> | </div> | ||
− | <p> Thanks to Prof. <b>Binguang Ma</b> for instructing our modeling. | + | <p> Thanks to Prof. <b>Binguang Ma</b> as our Primary PI and for supervising the whole project and instructing our modeling. |
</p> | </p> | ||
<p> Thanks to Prof. <b>Jin He</b> as our Secondary PI and Prof. <b>Shan Li</b> for instructing our | <p> Thanks to Prof. <b>Jin He</b> as our Secondary PI and Prof. <b>Shan Li</b> for instructing our | ||
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for giving us advices. | for giving us advices. | ||
</p> | </p> | ||
+ | <p> Thanks to <b>Weitong Guo</b> from iGEM-2017, Huazhong Agricultural University, for the training of | ||
+ | new dry-labers during winter vacation. | ||
<p style="margin-bottom:30px"> We really appreciate your support! | <p style="margin-bottom:30px"> We really appreciate your support! | ||
</p> | </p> |
Latest revision as of 02:00, 18 October 2018
Our project was designed by Zhujun Xia and all circuits were designed by Zhujun Xia, YinQing Zeng and Mo Qiqin.
sifA was knocked out by Zhujun Xia, Zhuoqi Huang and Wenxin Hu also gave assistance.
Intracellular Environment-Dependent Circuits:
The intracellular environment-dependent circuits were designed by Mo Qiqin and Zhujun Xia.
The intracellular environment-dependent circuits were constructed by Mo Qiqin. Ruonan Tian also gave assistance.
Chemical Control System:
The ATc induction chemical control system was constructed by He Yang and Zhuoqi Huang also gave assistance.
The TetR-eGFP fusion protein expression system for TetR expression measurement was constructed by He Yang and Zhuoqi Huang also gave assistance.
Targeting Specificity and Surface-Displaying:
RGD-inserted OmpA circuit clone and construction of Salmonella SL1344_RS05255 (OmpA coding sequence) were completed by YinQing Zeng and Lingyu Zhong.
The circuit of surface-display system was constructed by Lingyu Zhong. Optimation of OmpA in E. Coli was completed by Zhujun Xia.
Plasmid Standardization:
Plasmid standardization was completed by Zhiqing Guo, Hangxi Fu and Zhuoqi Huang by using 3A Assembly.
Plasmid Submission:
Submitted plasmids (Pcs2-eGFP-GSDMD FL, Pcs2-eGFP-GSDMD-N275 and two GSDMD full length mutations) were constructed by Zhiqing Guo and Wenxin Hu.
The description of submitted plasmids by ATP assay and cell phenotype via transfection was designed and completed by Zhujun Xia.
Bacterial phenotype verifications were designed and completed by He Yang, including the OD measurement and CFU measurement of chemical control system.
Cell phenotype verifications were designed and completed by Zhujun Xia, including the verification of chemical control system, and targeting specificity, all the related circuits and the demonstration of the final work system.
Safety verification was designed and completed by Zhujun Xia, which demonstrated our project is safe enough to be used.
InterLab was conducted by Xichen Rao, Zhujun Xia, Ruonan Tian and Heng Heng also gave assistance. Our data passed the check of the Measurement Committee successfully.
Mathematical model and app about salmonella infection and GSDMD release were built by Songtao Chen.
ATc model which predicted the relationship between concentration of ATc and concentration of GSDMD was built by Xichen Rao.
The modeling on adjusting the strength of RBS and ATc with fixed GSDMD-N concentration was completed by Yini Miao.
The modeling of the promoter sifA and promoter ent was completed by Ruonan Tian.
The applet of "Bacterial Colony Counter" was designed and programmed by Shuguang Wang.
Collaboration and communication with other teams were completed by Yinqing Zeng and all the other team members.
Handicraft manufacture was designed and completed by Yinqing Zeng.
Questionnaire preparation, the advertisement of synthetic biology and preparation of popular science articles were completed by Heng Heng.
Two posters for Eurasian exchange meetings and CCIC were completed by Ruonan Tian.
Our wiki framework was designed and programmed by Shuguang Wang and our artist were Yuying Xiang and Kening Chen.
Our blog was built by Shuguang Wang.
The plate reader machine operation was conducted by Mo Qiqin.
Chief cleaner of the laboratory was Xichen Rao.
For each parts of experiments, our appreciations are as follow:
Thanks to Prof. Shan Li’s laboratory, Huazhong Agricultural University, for sharing the cell lines of HEK293T, HELA, MCF7, iBMDM, so that our experiment can process successfully.
Thanks to Dr. Feng Shao’s laboratory, for sharing the cell line of GSDMD-/- HELA cells, which provides us great support.
Thanks to Dr. Yi Zeng and Dr. Ms Hong Fan, Shanghai Institute of Biochemistry and Cell Biology, CAS, for sharing the cell line of MDA-MB-231.
Thanks to Prof. Shan Li’s laboratory, Huazhong Agricultural University, for sharing the strain of Salmonella enterica var. Typhimurium SL1344.
Thanks to Dr. Chenli Liu from SIAT CSynBER for generously sharing the material (Strain: E.coli K-12 MG1655 with mCherry in the genome).
Thanks to Prof. Shan Li’s laboratory, Huazhong Agricultural University, for providing the protocol and material of gene editing based on two-step allelic exchange, which provides us with great help.
Thanks to Prof. Shan Li’s laboratory, Huazhong Agricultural University, for sharing the plasmid backbone (Pcs2-eGFP) to us.
Thanks to GeneScript and IDT for DNA sequence synthesis service.
Thanks to SnapGene and MathWorks for free software supporting.
Thanks to College of Life Science and Technology and Huazhong Agricultural University for providing the working site and fund for us.
Thanks to Prof. Binguang Ma as our Primary PI and for supervising the whole project and instructing our modeling.
Thanks to Prof. Jin He as our Secondary PI and Prof. Shan Li for instructing our experiment.
Thanks to Prof. Jin He, Prof. Shan Li, Prof. Gang Cao, Prof. Donghai Peng and Prof. Ming Sun for giving us advices at the proposal presentation.
Thanks to Kening Chen, Wenqi Huang and Haimeng Li from iGEM-2017, Huazhong Agricultural University, for giving us advices.
Thanks to Weitong Guo from iGEM-2017, Huazhong Agricultural University, for the training of new dry-labers during winter vacation.
We really appreciate your support!