Difference between revisions of "Team:CPU CHINA/Improve"

 
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<h1>Human practice summary for silver medal
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<h1>Part improvement
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<h4>Hepatitis C virus nonstructural protein 5B (NS5B) is a RNA-dependent RNA polymerase (RdRp) which initiates de novo RNA synthesis with the help of specific RNA promoters. In our project, the RNA interference is conditional because the pri-miRNA analogue which partially pairs with an inhibitory strand is unable to be processed by DROSHA. Once NS5B is introduced to catalyze a substitutional hybridation to release the pri-miRNA in the nucleus, the analogue gets to evolve into miRNA and interferes with the target gene mRNA.
 
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<h4>Through series of visiting and interviews with professionals (learn the details below in integrated human practice summary), we were eventually able to apply our RNA interference strategy to HCC therapeutic. Then we continued our human practice by promoting the public awareness of HCC, our project and synthetic biology.
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<h4>Since the existent NS5B from Standard Parts collection which can not translocate into the nucleus does not suit our needs, we tend to design our own part that is able to do its job in the nucleus. After deep consideration, we decided to improve the function of the part submitted by iGEM14<u> </u>Warwick (<a href="http://parts.igem.org/Part:BBa_K1442028">BBa_K1442028</a>). We load a nuclear location sequence (NLS) to the N terminal of NS5B (NS5B<sup>NLS</sup>) to transport the RdRP into the nucleus (<a herf="http://parts.igem.org/Part:BBa_K2624002">BBa_K2624002</a>). We demonstrate this improvement by indirect immunofluorescence. As the following figure shows, NS5B with NLS (NLS+) have a higher ability of nuclear translocation than NS5B without NLS (NLS-).
 
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<h4>First, we were curious as to how people view HCC. So a public survey about HCC was conducted, and then we realized that their understanding about this deadly disease is generally shallow and misguided. In light of such situation we felt the need for a booklet which is now accessible online in which basic HCC prevention and treatment are discussed.
 
 
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<h4>Then, to widen the impact of our project on campus, we organized an Innovation Experiment for College Students study group based on our RNAi system and three junior students joined to learn our project. They were able to practice some experimental techniques such as molecular cloning.
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<h4>Promoting synthetic biology has always been our mission, this iGEM season marks the third year of our life science student club CPU Bio-X Club. The popularity of this club elevated greatly this year with weekly professional lectures and a big campus-ranged event. We also tried some other novel ways including the TED<sup>x</sup> speech “<i>Synthetic biology, creating a better future</i>” and on-street interviews.
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<h4>This year, CPU-CHINA conducted integrated human practices by link it with the design of our project, not only in parts design, but also in our future business plan since we were eager to develop a therapy that suits public needs. In the first place, we developed a highly selective novel RNA interference strategy and hoped that it could be applied to certain diseases. We interviewed with clinical oncologists, researched on relevant literature and decided on HCC. We also added disease-specific promoters which could help reduce side effects usually involved in cancer therapy, as well as a small-molecule-dependent switch which would be useful for individual administration through our interview and literature researching as well.  
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<h4>Since the specific parts had been designed, testing the practical plausibility of our application became the priority. We interviewed with several business experts to discuss the project in real business settings, which led to a business plan. In that plan we thoroughly considered the current market, our competition and our strength.
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<h4>Last, we tried to learn patients’ reflection about our strategy. We once thought they might feel reluctant because gene therapy is not the most common method for cancer treatment, but soon we found that such concern was unnecessary. We communicated with patients from a huge cancer association, especially one that has much benefitted from novel cancer therapy. He confirmed the significance of specific and effective drugs regardless of how anything is popular at the moment.
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Latest revision as of 14:11, 6 December 2018

Part improvement

Hepatitis C virus nonstructural protein 5B (NS5B) is a RNA-dependent RNA polymerase (RdRp) which initiates de novo RNA synthesis with the help of specific RNA promoters. In our project, the RNA interference is conditional because the pri-miRNA analogue which partially pairs with an inhibitory strand is unable to be processed by DROSHA. Once NS5B is introduced to catalyze a substitutional hybridation to release the pri-miRNA in the nucleus, the analogue gets to evolve into miRNA and interferes with the target gene mRNA.

Since the existent NS5B from Standard Parts collection which can not translocate into the nucleus does not suit our needs, we tend to design our own part that is able to do its job in the nucleus. After deep consideration, we decided to improve the function of the part submitted by iGEM14 Warwick (BBa_K1442028). We load a nuclear location sequence (NLS) to the N terminal of NS5B (NS5BNLS) to transport the RdRP into the nucleus (BBa_K2624002). We demonstrate this improvement by indirect immunofluorescence. As the following figure shows, NS5B with NLS (NLS+) have a higher ability of nuclear translocation than NS5B without NLS (NLS-).