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- /wiki/pages/engineering.html
- /wiki/menu.html
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wiki/menu.html
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<a href="#" style="cursor:default;">Project</a> <a href="#" style="cursor:default;">Project</a>
<ul class="son"> <ul class="son">
<li><a href="{{ url_for('pages', page='description') }}">Description</a></li> <li><a href="{{ url_for('pages', page='description') }}">Description</a></li>
<li><a href="{{ url_for('pages', page='engineering') }}">Engineer Success</a></li> <li><a href="{{ url_for('pages', page='engineering') }}">Engineering</a></li>
<li><a href="{{ url_for('pages', page='proof-of-concept') }}">Proof Of concept</a></li>
<li><a href="{{ url_for('pages', page='safety') }}">Safety</a></li> <li><a href="{{ url_for('pages', page='safety') }}">Safety</a></li>
<li><a href="{{ url_for('pages', page='contribution') }}">Contribution</a></li> <li><a href="{{ url_for('pages', page='contribution') }}">Contribution</a></li>
</ul> </ul>
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wiki/pages/engineering.html
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margin-left: 130px; margin-left: 130px;
} }
</style> </style>
</head> </head>
<body> <body>
{% extends "layout.html" %} {% extends "layout.html" %}
{% block title %}Engineering Success{% endblock %} {% block title %}Engineering{% endblock %}
{% block page_content %} {% block page_content %}
<div class="sidebar"> <div class="sidebar">
<ul> <ul>
<li><a href="#Introduction">Introduction</a></li> <li><a href="#description">General Description</a></li>
<li><a href="#Muscone molecular switch">Muscone molecular switch</a></li> <li><a href="#topic1">Topic1</a></li>
<li><a href="#Colonization system">Colonization system</a></li> <li><a href="#topic2">Topic2</a></li>
<li><a href="#Inhalation of muscone">Inhalation of muscone</a></li> <li><a href="#topic3">Topic3</a></li>
<li><a href="#topic4">Topic4</a></li>
</ul> </ul>
</div> </div>
...@@ -52,108 +54,61 @@ ...@@ -52,108 +54,61 @@
<div class="row mt-4"> <div class="row mt-4">
<div class="col-lg-12"> <div class="col-lg-12">
<h2 id="Introduction"> <h2 id="description">
<h2>Introduction</h2> <h2>General Description</h2>
<hr>
<p>The core of our treatment project revolves around three aspects: </p>
<p>Effectiveness: The therapy system we designed can effectively induce yeast to secrete lactic acid; </p>
<p>Safety: Our engineered yeast can specifically colonize the patient's lesion site and die after leaving the patient's body environment; </p>
<p>Feasibility: Muscone can reach the intestines at a higher concentration through inhalation.</p>
<p>We designed detailed physical and chemical experiments to verify the first two aspects; for the third point, due to the safety restrictions imposed by iGEM on participating teams, we only used theoretical models such as gas molecule diffusion to illustrate it. Unfortunately, the plasmid expression system in yeast is not very stable. In addition, the molecular switch of muscone we designed has a complex interplay with the existing signaling pathways in yeast cells.</p>
<p>Therefore, validating experiments is very difficult, especially considering the time constraints of the iGEM competition. Despite referencing a large amount of papers and materials, most of the data we obtained is still frustrating. It is also very difficult to explain these anomalous data. We have overcome all these obstacles and successfully established a more mature muscone molecular switch and colonization system in the yeast system. This indicates that the concept of inhalation therapy with muscone is a theoretically feasible one. </p>
<p>The muscone molecular switch we introduced from the yeast system is a very novel, simple, and responsive regulatory method. By modifying the downstream response signals, it can be easily applied to the design of other genetic engineering projects.</p>
</div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h2 id="Muscone molecular switch">
<h2>Muscone molecular switch</h2>
<hr> <hr>
<div class="image-container" style="display: flex; flex-direction: column; align-items: center;"> <p>Inflammatory Bowel Disease (IBD) is an indeterminate colitis and autoimmune inflammatory bowel disease that involves the ileum, rectum, and colon. Due to its unknown pathogenesis, it is difficult to cure and prone to relapse, which is then referred to as the “green cancer”. As the incidence of IBD is increasing globally, the poor therapeutic experience, which the patients are required to receive long-term hormone injection and biologic combination therapy, is determined to seek a breakthrough. Our team stands to develop a more favorable treatment experience. Relevant studies have shown that lactic acid has an inhibitory effect on abnormally activated dendritic cells and cytotoxic T-cells in autoimmune diseases, and thus is expected to improve the symptoms of IBD patients. Based on the research, we designed a yeast engineered bacterium. We introduced a muscone detecting switch in the engineered yeast and activated the downstream pathway to express lactate dehydrogenase to produce lactic acid. Ultimately, the engineered bacteria will colonize the small intestine, enabling the patient to treat IBD by inhaling muscone gas in vitro therefore induces the yeast to synthesize a low amount of stabilized lactic acid in vivo. Aroma therapy has the potential to be a new trend in long-term treatment for IBD as it can reduce the discomfort of treatment and improve the long-term treatment experience. </p>
<img src="https://static.igem.wiki/teams/5187/wiki-engineer-fig/figure1.jpg" alt="ibd_figure" class="shadowed-image" style="width: 45%; max-width: 400px;">
<p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 1 Muscone molecular switch-regulated lactic acid secretion system</p>
</div>
</div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h3>Yeast can normally secrete lactic acid</h3>
<p>We introduced the lactate dehydrogenase gene (ldhA) regulated by a galactose promoter into yeast cells, induced it under different conditions, and measured the lactic acid content in the supernatant. We found that, aside from a small amount of leakage expression of the ldhA gene itself, galactose can very efficiently induce yeast to secrete sufficient lactic acid into the external environment. It is worth noting that because glucose is a better carbon source for yeast compared to galactose, the difference in carbon sources can have some effects on lactic acid secretion beyond just gene expression, which we have not demonstrated here. There is a more detailed discussion in the wet lab section.</p>
<div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
<img src="https://static.igem.wiki/teams/5187/wiki-engineer-fig/figure2.png" alt="ibd_figure" class="shadowed-image" style="width: 30%; max-width: 300px;">
<p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 2 Results of yeast-induced lactic acid secretion. This image shows the differences in lactic acid secretion by yeast induced by different carbon sources (glucose or galactose).(wt: wild-type yeast, ldhA: yeast transformed with a plasmid containing the ldhA gene)</p>
</div> </div>
<div class="image-container">
<img src="https://static.igem.wiki/teams/5187/figure/ibd-figure.jpg" alt="ibd_figure" class="shadowed-image">
</div> </div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h3>The muscone molecular switch can function efficiently</h3>
<p>We simultaneously introduced plasmid vectors expressing the muscone receptor and the corresponding Gα protein into yeast cells, along with a plasmid vector expressing the GFP protein controlled by the pFUS1 promoter. We used galactose to induce the expression of receptors and Gα proteins, and added a certain concentration of muscone for induction. The results are as follows. We found that, aside from the weak background expression, muscone can very efficiently induce the expression of the downstream controlled GFP gene.</p>
<div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
<img src="https://static.igem.wiki/teams/5187/wiki-engineer-fig/figure3.png" alt="ibd_figure" class="shadowed-image" style="width: 45%; max-width: 400px;">
<p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 3 Yeast specifically expresses GFP through designed pathways in the condition of muscone inducing</p>
</div>
</div> </div>
</div>
<div class="row mt-4">
<div class="row mt-4"> <div class="col-lg-12">
<div class="col-lg-12"> <h2 id="topic1">
<h3>The muscone molecular switch can control lactic acid secretion in a concentration-dependent manner</h3> <h2>Topic1</h2>
<p>We combined the muscone molecular switch with the lactate dehydrogenase gene to measure the differences in the rate of lactic acid secretion by yeast induced by different concentrations of muscone.</p> <hr>
<p>Tsinghua University is a public university in Haidian, Beijing. It is affiliated with and funded by the Ministry of Education of China. The university is part of Project 211, Project 985, and the Double First-Class Construction. It is also a member of the C9 League. Tsinghua University's campus is situated in northwest Beijing, on the site of the former imperial gardens of the Qing dynasty. Currently, the university has 21 schools and 59 departments, with faculties in science, engineering, humanities, law, medicine, history, philosophy, economics, management, education and art.</p>
<p>Notable alumni who have held senior positions in Chinese politics include current general secretary and president of China, Xi Jinping, former general secretary and president of China Hu Jintao, former chairman of the National People's Congress Wu Bangguo, former premier Zhu Rongji, and the former first vice premier Huang Ju. This also includes politicians like Wu Guanzheng, former governor of the People's Bank of China Zhou Xiaochuan, former minister of finance Lou Jiwei, general Sun Li-jen, Liang Qichao, and more. Since 2016, Tsinghua graduates who have political prominence are disproportionately greater in number than graduates of other famous universities.</p>
</div>
</div> </div>
</div>
<div class="row mt-4">
<div class="row mt-4"> <div class="col-lg-12">
<div class="col-lg-12"> <h2 id="topic2">
<h2 id="Colonization system"> <h2>Topic2</h2>
<h2>Colonization system</h2>
<hr> <hr>
<div class="image-container" style="display: flex; flex-direction: column; align-items: center;"> <p>Tsinghua University engages in extensive research and offers 51 bachelor's degree programs, 139 master's degree programs, and 107 doctoral programs through 20 colleges and 57 departments covering a broad range of subjects, including science, engineering, arts and literature, social sciences, law, medicine. Along with its membership in the C9 League, Tsinghua University affiliations include the Association of Pacific Rim Universities, a group of 50 leading Asian and American universities, Washington University in St. Louis's McDonnell International Scholars Academy, a group of 35 premier global universities, and the Association of East Asian Research Universities, a 17-member research collaboration network of top regional institutions. Tsinghua is an associate member of the Consortium Linking Universities of Science and Technology for Education and Research (CLUSTER). Tsinghua is a member of a Low Carbon Energy University Alliance (LCEUA), together with the University of Cambridge and the Massachusetts Institute of Technology (MIT).</pp>
<img src="https://static.igem.wiki/teams/5187/wiki-engineer-fig/figure4.png" alt="ibd_figure" class="shadowed-image" style="width: 45%; max-width: 400px;"> <p>School of Life Sciences was first established in 1926 under the name Department of Biology. Botanist Qian Chongshu took up the first dean.During the nationwide reorganization of universities in the early 1950s, the Department of Biology was merged into other universities, namely Peking University etc., resulting in a vacancy in the field of biological research in Tsinghua for almost 30 years.In June 1984, decisions were made about the reestablishment of the Department of Biology, and the department officially reopened in September. During the reestablishment the Department of Biology of Peking University, the Institute of Biophysics of Chinese Academy of Sciences, and many other institutes as well as biologists provided valuable support and help. The department changed its name to the current name in September 2009. As of 2013, structural biologist and foreign associate of National Academy of Sciences of United States Dr. Wang Hongwei (王宏伟) is the current dean of School of Life Sciences. The school currently has 129 professors and employees, around 600 undergraduates (including the candidates of Tsinghua University – Peking Union Medical College joint MD program).</p>
<p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 4 Thiosulfate-induced Als3-dependent colonization system</p>
</div> </div>
</div> </div>
</div>
<div class="row mt-4">
<div class="row mt-4"> <div class="col-lg-12">
<div class="col-lg-12"> <h2 id="topic3">
<h3>The Als3 protein can effectively adhere yeast to human intestinal epithelium</h3> <h2>Topic3</h2>
<p>We introduced the Als3 gene, regulated by a galactose promoter, into yeast and found that after the addition of galactose, it could attach in greater numbers to human intestinal epithelium compared to wild-type yeast.</p> <hr>
</div> <p>Admission to Tsinghua for both undergraduate and graduate schools is extremely competitive. Undergraduate admissions for domestic students is decided through the gaokao, the Chinese national college entrance exam, which allows students to list Tsinghua University among their preferred college choices. While selectivity varies by province, the sheer number of high school students applying for college each year has resulted in overall acceptance rates far lower than 0.1% of all test takers. Admission to Tsinghua's graduate schools is also very competitive. Only about 16% of MBA applicants are admitted each year.</p>
</div> <p>Department of Mathematical Sciences
The Department of Mathematical Sciences (DMS) was established in 1927.
<div class="row mt-4"> In 1952, Tsinghua DMS was merged with the Peking University Department of Mathematical Sciences. Then in 1979 it was renamed "Department of Applied Mathematics", and renamed again in 1999 to its current title.
<div class="col-lg-12"> Tsinghua DMS has three institutes at present, the institute of Pure Mathematics which has 27 faculty members, the Institute of Applied Mathematics and Probability and Statistics which has 27 faculty members, and the Institute of Computational Mathematics and Operations Research which has 20 faculty members. There are currently about 400 undergraduate students and 200 graduate students.</p>
<h3>Thiosulfate receptors can work effectively</h3> </div>
<p>We introduced plasmids expressing downstream regulated GFP and thiosulfate receptors into yeast simultaneously and found that thiosulfate can significantly enhance the intensity and proportion of GFP signal expression in yeast.</p>
</div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h3>Thiosulfate receptors can work effectively</h3>
<p>……to be continue</p>
</div> </div>
</div>
<div class="row mt-4">
<div class="row mt-4"> <div class="col-lg-12">
<div class="col-lg-12"> <h2 id="topic4">
<h2 id="Inhalation of muscone"> <h2>Topic4</h2>
<h2>Inhalation of muscone</h2>
<hr> <hr>
<p>Tsinghua University is consistently ranked among the top universities in the Asia-Pacific according to major international university rankings. Tsinghua University ranked No. 1 in China, the whole of Asia-Oceania region and emerging countries according to the Times Higher Education, with its industry income, research, and teaching performance indicator placed at 1st, 4th and 9th respectively in the world. Internationally, Tsinghua was regarded as the most reputable Chinese university by the Times Higher Education World Reputation Rankings where, it has ranked 8th globally and 1st in the Asia-Pacific.</p>
<p>The Engineering Research Center for Navigation Technology is a relatively young institute in the Department of Precision Instrument which was established in 2000, with the intention to "[pursue] excellence in the research and development in the field of high-accuracy inertial instruments and navigation technology, as well as in MEMS inertial sensor fields, and to provide advanced training for future scientists and engineers in the field of inertial technology." Its research interests cover high-accuracy inertial instruments and navigation technology, MEMS inertial sensors and systems, and precise electro-mechanical control systems and their application. As of 2012, the area of the center is 2900 square meters, including approximately 550 square meters of clean rooms. Equipment and instruments in this center are worth over 50 million RMB (US$7.56 million).</p>
</div>
<div class="image-container">
<img src="https://static.igem.wiki/teams/5187/figure/ibd-figure.jpg" alt="ibd_figure" class="shadowed-image">
</div> </div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h3>Thiosulfate receptors can work effectively</h3>
<p>……to be continue</p>
</div> </div>
</div>
</body> </body>
</html> </html>
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wiki/pages/proof-of-concept.html 0 → 100644
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<!DOCTYPE html>
<html lang="en">
<head>
<meta charset="UTF-8">
<meta name="viewport" content="width=device-width, initial-scale=1.0">
<link rel="icon" type="image/png" href="https://static.igem.wiki/teams/5187/art/icon1.png" sizes="364x370">
<title>Tsinghua - IGEM 2024</title>
<style>
body {
font-family: Arial, sans-serif;
line-height: 1.6;
margin: 0;
padding: 0;
}
.content {
padding: 20px;
max-width: 800px;
margin: 0 auto;
}
h2 {
scroll-margin-top: 60px;
}
.row.mt-4 {
margin-right: 100px;
margin-left: 130px;
}
</style>
</head>
<body>
{% extends "layout.html" %}
{% block title %}Proof Of Concept{% endblock %}
{% block page_content %}
<div class="sidebar">
<ul>
<li><a href="#Introduction">Introduction</a></li>
<li><a href="#Muscone molecular switch">Muscone molecular switch</a></li>
<li><a href="#Colonization system">Colonization system</a></li>
<li><a href="#Inhalation of muscone">Inhalation of muscone</a></li>
</ul>
</div>
<div class="progress-container">
<svg class="progress-bar-circle" width="60" height="60">
<circle class="progress-circle" cx="30" cy="30" r="25" stroke-width="5" fill="transparent"></circle>
</svg>
<div class="progress-text">0%</div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h2 id="Introduction">
<h2>Introduction</h2>
<hr>
<p>The core of our treatment project revolves around three aspects: </p>
<p>Effectiveness: The therapy system we designed can effectively induce yeast to secrete lactic acid; </p>
<p>Safety: Our engineered yeast can specifically colonize the patient's lesion site and die after leaving the patient's body environment; </p>
<p>Feasibility: Muscone can reach the intestines at a higher concentration through inhalation.</p>
<p>We designed detailed physical and chemical experiments to verify the first two aspects; for the third point, due to the safety restrictions imposed by iGEM on participating teams, we only used theoretical models such as gas molecule diffusion to illustrate it. Unfortunately, the plasmid expression system in yeast is not very stable. In addition, the molecular switch of muscone we designed has a complex interplay with the existing signaling pathways in yeast cells.</p>
<p>Therefore, validating experiments is very difficult, especially considering the time constraints of the iGEM competition. Despite referencing a large amount of papers and materials, most of the data we obtained is still frustrating. It is also very difficult to explain these anomalous data. We have overcome all these obstacles and successfully established a more mature muscone molecular switch and colonization system in the yeast system. This indicates that the concept of inhalation therapy with muscone is a theoretically feasible one. </p>
<p>The muscone molecular switch we introduced from the yeast system is a very novel, simple, and responsive regulatory method. By modifying the downstream response signals, it can be easily applied to the design of other genetic engineering projects.</p>
</div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h2 id="Muscone molecular switch">
<h2>Muscone molecular switch</h2>
<hr>
<div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
<img src="https://static.igem.wiki/teams/5187/wiki-engineer-fig/figure1.jpg" alt="ibd_figure" class="shadowed-image" style="width: 45%; max-width: 400px;">
<p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 1 Muscone molecular switch-regulated lactic acid secretion system</p>
</div>
</div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h3>Yeast can normally secrete lactic acid</h3>
<p>We introduced the lactate dehydrogenase gene (ldhA) regulated by a galactose promoter into yeast cells, induced it under different conditions, and measured the lactic acid content in the supernatant. We found that, aside from a small amount of leakage expression of the ldhA gene itself, galactose can very efficiently induce yeast to secrete sufficient lactic acid into the external environment. It is worth noting that because glucose is a better carbon source for yeast compared to galactose, the difference in carbon sources can have some effects on lactic acid secretion beyond just gene expression, which we have not demonstrated here. There is a more detailed discussion in the wet lab section.</p>
<div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
<img src="https://static.igem.wiki/teams/5187/wiki-engineer-fig/figure2.png" alt="ibd_figure" class="shadowed-image" style="width: 30%; max-width: 300px;">
<p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 2 Results of yeast-induced lactic acid secretion. This image shows the differences in lactic acid secretion by yeast induced by different carbon sources (glucose or galactose).(wt: wild-type yeast, ldhA: yeast transformed with a plasmid containing the ldhA gene)</p>
</div>
</div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h3>The muscone molecular switch can function efficiently</h3>
<p>We simultaneously introduced plasmid vectors expressing the muscone receptor and the corresponding Gα protein into yeast cells, along with a plasmid vector expressing the GFP protein controlled by the pFUS1 promoter. We used galactose to induce the expression of receptors and Gα proteins, and added a certain concentration of muscone for induction. The results are as follows. We found that, aside from the weak background expression, muscone can very efficiently induce the expression of the downstream controlled GFP gene.</p>
<div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
<img src="https://static.igem.wiki/teams/5187/wiki-engineer-fig/figure3.png" alt="ibd_figure" class="shadowed-image" style="width: 45%; max-width: 400px;">
<p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 3 Yeast specifically expresses GFP through designed pathways in the condition of muscone inducing</p>
</div>
</div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h3>The muscone molecular switch can control lactic acid secretion in a concentration-dependent manner</h3>
<p>We combined the muscone molecular switch with the lactate dehydrogenase gene to measure the differences in the rate of lactic acid secretion by yeast induced by different concentrations of muscone.</p>
</div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h2 id="Colonization system">
<h2>Colonization system</h2>
<hr>
<div class="image-container" style="display: flex; flex-direction: column; align-items: center;">
<img src="https://static.igem.wiki/teams/5187/wiki-engineer-fig/figure4.png" alt="ibd_figure" class="shadowed-image" style="width: 45%; max-width: 400px;">
<p style="text-align: center; font-size: 0.9em; margin-top: 10px;">fig 4 Thiosulfate-induced Als3-dependent colonization system</p>
</div>
</div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h3>The Als3 protein can effectively adhere yeast to human intestinal epithelium</h3>
<p>We introduced the Als3 gene, regulated by a galactose promoter, into yeast and found that after the addition of galactose, it could attach in greater numbers to human intestinal epithelium compared to wild-type yeast.</p>
</div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h3>Thiosulfate receptors can work effectively</h3>
<p>We introduced plasmids expressing downstream regulated GFP and thiosulfate receptors into yeast simultaneously and found that thiosulfate can significantly enhance the intensity and proportion of GFP signal expression in yeast.</p>
</div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h3>Thiosulfate receptors can work effectively</h3>
<p>……to be continue</p>
</div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h2 id="Inhalation of muscone">
<h2>Inhalation of muscone</h2>
<hr>
</div>
</div>
<div class="row mt-4">
<div class="col-lg-12">
<h3>Thiosulfate receptors can work effectively</h3>
<p>……to be continue</p>
</div>
</div>
</body>
</html>
{% endblock %}
\ No newline at end of file
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