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| − | + | <ul id="left-nav"> | |
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<li> | <li> | ||
<a>PROJECT</a> | <a>PROJECT</a> | ||
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<li><a href="https://2018.igem.org/Team:BIT-China/ExperimentsFeedback">Feedback</a></li> | <li><a href="https://2018.igem.org/Team:BIT-China/ExperimentsFeedback">Feedback</a></li> | ||
<li><a href="https://2018.igem.org/Team:BIT-China/ExperimentsOutput">Output</a></li> | <li><a href="https://2018.igem.org/Team:BIT-China/ExperimentsOutput">Output</a></li> | ||
| − | <li><a href="https://2018.igem.org/Team:BIT-China/ | + | <li><a href="https://2018.igem.org/Team:BIT-China/Results">Results</a></li> |
</ul> | </ul> | ||
</li> | </li> | ||
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<ul> | <ul> | ||
<li><a href="https://2018.igem.org/Team:BIT-China/Model">Overview</a></li> | <li><a href="https://2018.igem.org/Team:BIT-China/Model">Overview</a></li> | ||
| − | <li><a href="https://2018.igem.org/Team:BIT-China/FluorescentProbesModel">Fluorescent | + | <li><a href="https://2018.igem.org/Team:BIT-China/FluorescentProbesModel">Fluorescent Probe Model </a></li> |
| − | + | <li><a href="https://2018.igem.org/Team:BIT-China/H2O2DecompositionModel">H<sub>2</sub>O<sub>2</sub> | |
Decomposition Model</a></li> | Decomposition Model</a></li> | ||
| − | + | ||
<li><a href="https://2018.igem.org/Team:BIT-China/roGFP2-Orp1MichaelisEquationModel">roGFP2-Orp1 | <li><a href="https://2018.igem.org/Team:BIT-China/roGFP2-Orp1MichaelisEquationModel">roGFP2-Orp1 | ||
| − | Michaelis | + | Michaelis equation Model</a></li> |
</ul> | </ul> | ||
</li> | </li> | ||
<li> | <li> | ||
| − | <a> | + | <a>HUMAN PRACTICES</a> |
<ul> | <ul> | ||
| + | <li><a href="https://2018.igem.org/Team:BIT-China/HPOverview">Overview</a></li> | ||
<li><a href="https://2018.igem.org/Team:BIT-China/Human_Practices">Integrated Human Practices</a></li> | <li><a href="https://2018.igem.org/Team:BIT-China/Human_Practices">Integrated Human Practices</a></li> | ||
<li><a href="https://2018.igem.org/Team:BIT-China/Public_Engagement">Education & Public Engagement</a></li> | <li><a href="https://2018.igem.org/Team:BIT-China/Public_Engagement">Education & Public Engagement</a></li> | ||
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</li> | </li> | ||
</ul> | </ul> | ||
| − | + | <a href="https://2018.igem.org/Team:BIT-China"><img id="imgA" class="imgA-new-pos" src="https://static.igem.org/mediawiki/2018/4/46/T--BIT-China--iGEM2018-A_img.png " /></a> | |
| − | <a href="https://2018.igem.org/Team:BIT-China"><img id="imgA" class="imgA-new-pos" src=" | + | |
<!-- end --> | <!-- end --> | ||
<div class="PRO-white-head"></div> | <div class="PRO-white-head"></div> | ||
| − | < | + | <img class="PRO-BG" src="https://static.igem.org/mediawiki/2018/7/74/T--BIT-China--iGEM2018-project-bg.png"> |
| − | + | ||
| − | + | ||
| − | + | ||
| − | + | ||
<div id="PRO-content-all" class="content_container" style="margin-top:25vh;"> | <div id="PRO-content-all" class="content_container" style="margin-top:25vh;"> | ||
| + | <div style="font-family: 'kg_second_chances_solidRg';font-size: 30px;color: #131313;width: 100%;float:right;"> | ||
| + | <a style="border-bottom: 3px solid #131313;color: #131313;text-decoration: none;right: 0;float: right;">DESCRIPTION</a> | ||
| + | </div> | ||
<div id="PRO1" class="cd-section"> | <div id="PRO1" class="cd-section"> | ||
<div class="PRO-title-2"> | <div class="PRO-title-2"> | ||
| − | <a>Overview</a> | + | <a style="text-decoration:none;color:#131313;">Overview</a> |
</div> | </div> | ||
| − | <div class="PRO-content-all"> | + | <div class="PRO-content-all PRO-margin-toTitle2"> |
<div class="PRO-content"> | <div class="PRO-content"> | ||
<p class="PRO-content-p"> | <p class="PRO-content-p"> | ||
| − | + | To achieve our goal of detecting the antioxidants capacity of different products in living | |
| + | cells, it our system can be divided into three parts, regulator, feedback and output. | ||
</p> | </p> | ||
</div> | </div> | ||
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</div> | </div> | ||
| − | <div id="PRO2" class="cd-section"> | + | <div id="PRO2" class="cd-section PRO-margin-Title2Up"> |
<div class="PRO-title-2"> | <div class="PRO-title-2"> | ||
| − | <a>Regulator</a> | + | <a style="text-decoration:none;color:#131313;">Regulator</a> |
</div> | </div> | ||
<div class="PRO-content-all"> | <div class="PRO-content-all"> | ||
<div class="PRO-content"> | <div class="PRO-content"> | ||
| − | <p class="PRO-content-p"> | + | <p class="PRO-content-p PRO-margin-toTitle2"> |
Firstly, we choose to increase the intracellular ROS content, considering that: | Firstly, we choose to increase the intracellular ROS content, considering that: | ||
</p> | </p> | ||
| − | <ul class="PRO-content-p"> | + | <ul class="PRO-content-p PRO-margin-toContentP"> |
<li> | <li> | ||
● The level of ROS in normal cells is low and hard to detection. | ● The level of ROS in normal cells is low and hard to detection. | ||
</li> | </li> | ||
<li> | <li> | ||
| − | ● Make the cell under oxidative stress to simulate the normal cellular oxidative process or, rather, the aging process. | + | ● Make the cell under oxidative stress to simulate the normal cellular oxidative process |
| + | or, rather, the aging process. | ||
</li> | </li> | ||
<li> | <li> | ||
| − | ● The most intuitive effect of antioxidants is their reduction of intracellular ROS content. We can evaluate antioxidants indirectly through the remaining amount of ROS. | + | ● The most intuitive effect of antioxidants is their reduction of intracellular ROS |
| + | content. We can evaluate antioxidants indirectly through the remaining amount of ROS. | ||
</li> | </li> | ||
<li> | <li> | ||
| − | ● After reviewing most cell-based antioxidant detection methods recorded in literature, we discover that almost all protocols will induce cell in an oxidant stress firstly. | + | ● After reviewing most cell-based antioxidant detection methods recorded in literature, we |
| + | discover that almost all protocols will induce cell in an oxidant stress firstly. | ||
</li> | </li> | ||
</ul> | </ul> | ||
| − | <p class="PRO-content-p"> | + | <p class="PRO-content-p PRO-margin-toContentP"> |
| − | Therefore, we established this part to do such kind of thing. Meanwhile, we hope the way to stimulate the accumulation of ROS is easy and equipment-independent, which makes our system more available for customers. | + | Therefore, we established this part to do such kind of thing. Meanwhile, we hope the way to |
| + | stimulate the accumulation of ROS is easy and equipment-independent, which makes our system | ||
| + | more available for customers. | ||
</p> | </p> | ||
| − | <p class="PRO-content-p"> | + | <p class="PRO-content-p PRO-margin-toContentP"> |
| − | However, higher oxidative stress is always accompanied by lower survival rate, in hence we also need to do something to make our host cell more robust and prevent cells from dying for oxidation. | + | However, higher oxidative stress is always accompanied by lower survival rate, in hence we also |
| + | need to do something to make our host cell more robust and prevent cells from dying for | ||
| + | oxidation. | ||
</p> | </p> | ||
</div> | </div> | ||
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</div> | </div> | ||
| − | + | <div id="PRO2" class="cd-section PRO-margin-Title2Up"> | |
| − | <div id="PRO2" class="cd-section"> | + | |
<div class="PRO-title-2"> | <div class="PRO-title-2"> | ||
| − | <a>Feedback</a> | + | <a style="text-decoration:none;color:#131313;">Feedback</a> |
</div> | </div> | ||
<div class="PRO-content-all"> | <div class="PRO-content-all"> | ||
<div class="PRO-content"> | <div class="PRO-content"> | ||
| − | <p class="PRO-content-p"> | + | <p class="PRO-content-p PRO-margin-toTitle2"> |
| − | After increasing the concentration of ROS in cell, | + | After increasing the concentration of ROS in cell, there's still somethings we need to |
| + | consider: | ||
</p> | </p> | ||
| − | <ul class="PRO-content-p"> | + | <ul class="PRO-content-p PRO-margin-toContentP"> |
<li> | <li> | ||
● The high level of ROS will damage the cells and even lead to cells death. | ● The high level of ROS will damage the cells and even lead to cells death. | ||
</li> | </li> | ||
<li> | <li> | ||
| − | ● Low survival rate under high oxidant state may affect the signal output of our system, making our results not convincing | + | ● Low survival rate under high oxidant state may affect the signal output of our system, |
| + | making our results not convincing. | ||
</li> | </li> | ||
<li> | <li> | ||
| − | ● | + | ● It's hard to control the ROS accumulation in every assay, which means our test may be |
| + | NON-repeatable. | ||
</li> | </li> | ||
</ul> | </ul> | ||
| − | <p class="PRO-content-p"> | + | <p class="PRO-content-p PRO-margin-toContentP"> |
| − | Based on these reasons, we need to add an feedback system to control the concentration of ROS in vivo, make it higher than normal level, lower than death level to maintain cell viability and last, steady for test. We solve this problem by adding an ROS sensor, which has a suitable sensing threshold so that it can maintain the balancing between the high level of ROS in living cell and survival rate of cells. | + | Based on these reasons, we need to add an feedback system to control the concentration of ROS |
| + | in vivo, make it higher than normal level, lower than death level to maintain cell viability | ||
| + | and last, steady for test. We solve this problem by adding an ROS sensor, which has a suitable | ||
| + | sensing threshold so that it can maintain the balancing between the high level of ROS in living | ||
| + | cell and survival rate of cells. | ||
</p> | </p> | ||
</div> | </div> | ||
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</div> | </div> | ||
| − | <div id="PRO2" class="cd-section"> | + | <div id="PRO2" class="cd-section PRO-margin-Title2Up"> |
<div class="PRO-title-2"> | <div class="PRO-title-2"> | ||
| − | <a>Output</a> | + | <a style="text-decoration:none;color:#131313;">Output</a> |
</div> | </div> | ||
<div class="PRO-content-all"> | <div class="PRO-content-all"> | ||
<div class="PRO-content"> | <div class="PRO-content"> | ||
| − | <p class="PRO-content-p"> | + | <p class="PRO-content-p PRO-margin-toTitle2"> |
| − | Last but not least, the part of characterizing residual amount of intracellular ROS. | + | Last but not least, the part of characterizing residual amount of intracellular ROS. |
</p> | </p> | ||
| − | <p class="PRO-content-p"> | + | <p class="PRO-content-p PRO-margin-toContentP"> |
Different from chemical detecting methods, our signal output substance must be: | Different from chemical detecting methods, our signal output substance must be: | ||
</p> | </p> | ||
| − | <ul class="PRO-content-p"> | + | <ul class="PRO-content-p PRO-margin-toContentP"> |
<li> | <li> | ||
| − | ● simpler for users to detect | + | ● simpler for users to detect. |
</li> | </li> | ||
<li> | <li> | ||
| − | ● harmless to cells | + | ● harmless to cells. |
</li> | </li> | ||
<li> | <li> | ||
| − | ● accurate and sensitive enough to reflect the intracellular ROS content | + | ● accurate and sensitive enough to reflect the intracellular ROS content. |
</li> | </li> | ||
<li> | <li> | ||
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</ul> | </ul> | ||
| − | <p class="PRO-content-p"> | + | <p class="PRO-content-p PRO-margin-toContentP"> |
| − | To meet those needs, chemical redox probe has been out of our consideration, even though they have characteristic of extreme-high sensitivity, still they are irreversibility and unstable. We prefer a kind of characterization which can exist in living cell also in a consist level for stable measurement. | + | To meet those needs, chemical redox probe has been out of our consideration, even though they |
| + | have characteristic of extreme-high sensitivity, still they are irreversibility and unstable. | ||
| + | We prefer a kind of characterization which can exist in living cell also in a consist level for | ||
| + | stable measurement. | ||
</p> | </p> | ||
</div> | </div> | ||
</div> | </div> | ||
</div> | </div> | ||
| − | |||
| − | |||
</div> | </div> | ||
<!-- footer start --> | <!-- footer start --> | ||
| − | <div class="footer-all" style="margin-top: | + | <div class="footer-all" style="margin-top:65px;"> |
<div class="footer-left"> | <div class="footer-left"> | ||
| − | <img src=" | + | <img src="https://static.igem.org/mediawiki/2018/c/ca/T--BIT-China--iGEM2018-sponsor-1.png"> |
| − | + | <img src="https://static.igem.org/mediawiki/2018/0/07/T--BIT-China--iGEM2018-sponsor-2.png"> | |
| − | + | ||
| − | <img src=" | + | |
| − | + | ||
| − | + | ||
</div> | </div> | ||
<div class="footer-right"> | <div class="footer-right"> | ||
| − | <p style="font-family:'kg_second_chances_solidRg';font-size:12px;margin: 0;padding: 0 0 3px;">Contact</p> | + | <p style="font-family:'kg_second_chances_solidRg';font-size:12px;margin: 0;padding: 0 0 3px;margin-top: 8px;">Contact</p> |
| − | <p style="font-size:15px | + | <p style="font-size:15px;margin: 0;padding: 0;margin-top: 4px;">Institute of |
| − | Biotransformation and synthetic biosystem | + | Biotransformation and synthetic biosystem School of Chemistry and Chemical Engineering. |
| − | + | </p> | |
| − | + | <p style="font-size:15px;margin: 0;padding: 0;margin-top: 4px;"> | |
| − | + | Beijing Institute of Technology | |
| − | + | </p> | |
| + | <p style="font-size:15px;margin: 0;padding: 0;margin-top: 4px;"> | ||
| + | 100081, Beijing | ||
| + | </p> | ||
| + | <p style="font-size:15px;margin: 0;padding: 0;margin-top: 4px;"> | ||
| + | Email:lichun@bit.edu.cn | ||
| + | </p> | ||
| + | |||
| + | <p style="font-size:15px;margin: 0;padding: 0;margin-top: 4px;color: rgb(161, 161, 161);"> | ||
| + | Copyright © 2018 BIT-China | ||
</p> | </p> | ||
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Latest revision as of 00:24, 18 October 2018
To achieve our goal of detecting the antioxidants capacity of different products in living cells, it our system can be divided into three parts, regulator, feedback and output.
Firstly, we choose to increase the intracellular ROS content, considering that:
- ● The level of ROS in normal cells is low and hard to detection.
- ● Make the cell under oxidative stress to simulate the normal cellular oxidative process or, rather, the aging process.
- ● The most intuitive effect of antioxidants is their reduction of intracellular ROS content. We can evaluate antioxidants indirectly through the remaining amount of ROS.
- ● After reviewing most cell-based antioxidant detection methods recorded in literature, we discover that almost all protocols will induce cell in an oxidant stress firstly.
Therefore, we established this part to do such kind of thing. Meanwhile, we hope the way to stimulate the accumulation of ROS is easy and equipment-independent, which makes our system more available for customers.
However, higher oxidative stress is always accompanied by lower survival rate, in hence we also need to do something to make our host cell more robust and prevent cells from dying for oxidation.
After increasing the concentration of ROS in cell, there's still somethings we need to consider:
- ● The high level of ROS will damage the cells and even lead to cells death.
- ● Low survival rate under high oxidant state may affect the signal output of our system, making our results not convincing.
- ● It's hard to control the ROS accumulation in every assay, which means our test may be NON-repeatable.
Based on these reasons, we need to add an feedback system to control the concentration of ROS in vivo, make it higher than normal level, lower than death level to maintain cell viability and last, steady for test. We solve this problem by adding an ROS sensor, which has a suitable sensing threshold so that it can maintain the balancing between the high level of ROS in living cell and survival rate of cells.
Last but not least, the part of characterizing residual amount of intracellular ROS.
Different from chemical detecting methods, our signal output substance must be:
- ● simpler for users to detect.
- ● harmless to cells.
- ● accurate and sensitive enough to reflect the intracellular ROS content.
- ● Reversible so we can get real-time data.
To meet those needs, chemical redox probe has been out of our consideration, even though they have characteristic of extreme-high sensitivity, still they are irreversibility and unstable. We prefer a kind of characterization which can exist in living cell also in a consist level for stable measurement.
