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| <!-- 极速模式 --> | | <!-- 极速模式 --> |
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| <script type="text/javascript" src="https://2016.igem.org/Team:HUST-China/js/bootstrap?action=raw&ctype=text/javascript"></script> | | <script type="text/javascript" src="https://2016.igem.org/Team:HUST-China/js/bootstrap?action=raw&ctype=text/javascript"></script> |
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| html { | | html { |
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| box-sizing: content-box; | | box-sizing: content-box; |
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| #sideMenu .mainMenu_toggle, | | #sideMenu .mainMenu_toggle, |
| #sideMenu .mainMenu_Title, | | #sideMenu .mainMenu_Title, |
| #sideMenu .sideMenuTitle, | | #sideMenu .sideMenuTitle, |
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| <body> | | <body> |
| <header class="header"> | | <header class="header"> |
| + | <div class="navbar-logo"> |
| + | <a class="navbar-brand" href="https://igem.org/Main_Page"> |
| + | <img alt="Brand" src="https://2016.igem.org/wiki/skins/Igem/images/IGEM_white_letters.png"> |
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| + | <a class="navbar-brand" href="https://2016.igem.org/Team:HUST-China"> |
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| <nav class="navbar navbar-default"> | | <nav class="navbar navbar-default"> |
| <div class="container"> | | <div class="container"> |
− | <span class="menu"> Menu</span> | + | <span class="menu">Menu</span> |
| <div class="banner-top"> | | <div class="banner-top"> |
| <ul class="nav banner-nav"> | | <ul class="nav banner-nav"> |
− | <li><a class="active" href="#">HOME</a></li> | + | <li><a href="https://2016.igem.org/Team:HUST-China">HOME</a></li> |
− | <li class="dropdown1"><a class="down-scroll" href="#">PROJECT</a> | + | <li class="dropdown1"><a class="down-scroll active" href="#">PROJECT</a> |
| <ul class="dropdown2"> | | <ul class="dropdown2"> |
| <li><a href="https://2016.igem.org/Team:HUST-China/project/background">Background</a></li> | | <li><a href="https://2016.igem.org/Team:HUST-China/project/background">Background</a></li> |
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| </ul> | | </ul> |
| </li> | | </li> |
− | <li class="dropdown1"><a class="down-scroll" href="https://2016.igem.org/Team:HUST-China/Model">MODELING</a> | + | <li class="dropdown1"><a class="down-scroll" href="#">MODELING</a> |
− | <!-- <ul class="dropdown2">
| + | <ul class="dropdown2"> |
− | <li><a href="#">Mag-receiver</a></li> | + | <li><a href="https://2016.igem.org/Team:HUST-China/Model">Overview</a></li> |
− | <li><a href="#">Heating</a></li> | + | <li><a href="https://2016.igem.org/Team:HUST-China/Model/model-pro">Prokaryotic circuit</a></li> |
− | <li><a href="#">Thermo-regulator</a></li> | + | <li><a href="https://2016.igem.org/Team:HUST-China/Model/model-euk">Eukaryotic circuit</a></li> |
− | </ul> --> | + | <li><a href="https://2016.igem.org/Team:HUST-China/Model/model-app">Application circuit</a></li> |
| + | </ul> |
| </li> | | </li> |
− | <li class="dropdown1"><a class="down-scroll" href="">PARTS</a> | + | <li class="dropdown1"><a class="down-scroll" href="#">PARTS</a> |
| <ul class="dropdown2"> | | <ul class="dropdown2"> |
| <li><a href="https://2016.igem.org/Team:HUST-China/Parts">Summary</a></li> | | <li><a href="https://2016.igem.org/Team:HUST-China/Parts">Summary</a></li> |
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| </ul> | | </ul> |
| </li> | | </li> |
− | <li class="dropdown1"><a class="down-scroll" href="">TEAM</a> | + | <li class="dropdown1"><a class="down-scroll" href="#">TEAM</a> |
| <ul class="dropdown2"> | | <ul class="dropdown2"> |
| <li><a href="https://2016.igem.org/Team:HUST-China/Team">Team Roster</a></li> | | <li><a href="https://2016.igem.org/Team:HUST-China/Team">Team Roster</a></li> |
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| <div class="ref"></div> | | <div class="ref"></div> |
| <article> | | <article> |
| + | <p>iGEM is a stage for synthetic biology researchers to build a different world. Our team was inspired by previous iGEM teams and their projects. This year, we are eager to participate and contribute more.</p> |
| <!-- h2一级标题 --> | | <!-- h2一级标题 --> |
− | <h2>OVERVIEW</h2>
| |
| <!-- 文字 --> | | <!-- 文字 --> |
| <p> | | <p> |
− | This year, we want to offer a handy, adjustable and useful tool kit to everyone working on Synthetic Biology--Signal Filter. The core circuit is based on a positive feedback bi-stable or tri-stable system with the capacity of reducing noise and converting pulse signal into robust and persistent signal. To adapt to different experimental requirements,we also developed two versions of our Signal Filter--prokaryotic one and eukaryotic one. | + | <strong>At the beginning</strong>, we learned about previous projects from the websites, and we realized that, when dealing with real-world problems, issues like gene regulation, expression efficiency and system robustness are all important. When creating a great functional prototype, one should pay attention to all the details. And some of the new comers to synthetic biology may find it difficult to deal with. So we put it in our way: why not provide some validated gene expression regulation kits to iGEMers so that they no longer need to worry about building circuit, but focus on the key problem. |
| </p> | | </p> |
− | <!-- h3二级标题 -->
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− | <!-- <h3>SubHeading</h3>
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− | <p>dfwef</p>
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− | <h3>Hello world</h3>
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− | <p>fdwfewf</p> -->
| |
− | <h2>Prokaryotic version:</h2>
| |
| <p> | | <p> |
− | It is a tri-stable system adapted from bacteriophage λ operon. In the operon, promoter RE is activated by CII trascriptional activator. Ftsh is an ATP-dependent host metalloprotease which will normally degrade CII, while CIII serves as an inhibitor of Ftsh to free CII. And CI can function as a inhibitor to block pR, while Cro can bind pRE to stop the downstream gene’s expression. The positive feedback control under pRE is used to enhance pulse signal 1 and convert it into robust stable signal.
| + | We came up with different versions of gene expression circuits: lambda bacteriophage, kinases reaction pathway, ribo-switch, RNAi and so on. We made efforts on the circuit construction and valid characterization data while users mainly focus on the input and output which are highly concerned with real-world problems. In this manner, our kit can serve as a useful tool to save their time and energy. On the whole, our theme is to offer bricks to help others build their own project. |
| </p> | | </p> |
− | <!-- 图片 -->
| + | <p>And this summer we <strong>stepped a little forward</strong> to our goals. We worked out two versions of gene expression switch: a prokaryote tri-stable version derived from bacteriophage lambda and eukaryote bi-stable version based on ABA-response pathway. |
− | <img src="https://static.igem.org/mediawiki/2016/8/88/T--HUST-China--Description-Fig-Eukaryote.png" alt="" class="img-responsive">
| + | |
− | <p class="text-center">Fig*: circuit of prokaryote version serving as tri-stable signal filter</p> | + | |
− | <!-- <h2>Creation based on Signal Filter</h2> -->
| + | |
− | <!-- 文字 -->
| + | |
− | <p>
| + | |
− | When signal 1 comes,a double promoter structure will help induce Cro and CII downstream pRE. At the same time constitutively produced CIII will guarantee enough CII to enhance the transcription downstream the pRE. Thus,there forms a positive feedback loop to direct a fast and strong expression of cro. A quantity of Cro represses transcription under pRM by binding to Cro binding site and blocks gene of interest 2’s expression, so it turns to a stable state of expressing gene of interest 1.
| + | |
− | </p>
| + | |
− | <!-- <img src="" alt=""> -->
| + | |
− | <p>When signal 2 comes, under a certain inducible promoter,CI will be expressed and then binds to cI binding site within pR promoter,blocking gene of interest 1 and cIII’s expressions. With CIII’s reduction, FtsH gradually degrades CII to interrupt the stable state. Therefore, Cro’s expression immediately drop down , allowing transcription from pRM. The system turn to another stable state of gene of interest 2‘s expression.
| + | |
| </p> | | </p> |
| <p> | | <p> |
− | If there is no signal.Both genes of interest will be expressed. And if both of the two signal exist,the expression state will depend on the intensity of the initial signal input. | + | <strong>In the tri-stable gene expression switch,</strong> users should define two input and related output gene signals. The logic gate below can illustrate the circuit better: |
− | </p>
| + | |
− | <h2>Eukaryotic version:</h2>
| + | |
− | <p>It is a bi-stable system derived from Arabidopsis thaliana stress response system. Enzyme catalysis are core of this design to increase the efficiency of state transition.
| + | |
− | </p>
| + | |
− | <img src="https://static.igem.org/mediawiki/2016/8/88/T--HUST-China--Description-Fig-Eukaryote.png" alt="" class="img-responsive">
| + | |
− | <p class="text-center">Fig*: circuit of Eukaryote version serving as bii-stable signal filter</p>
| + | |
− | <p>Clade A protein phosphatases type 2C(PP2CA) and SUCROSE NONFERMENTING1-RELATED SUBFAMILY2(SnRK2s) protein kinase are both components of Abscisic acid signaling network in Arabidopsis thaliana. ABF2 is a leucine zipper transcription factor which basically binds ABA-response element (ABRE). ABF2 can be phosphorylated by SnRK2s and be efficiently dephosphorylated by PP2CA. Also, in the absence of Abscisic acid,SnRK2 kinases can be inactivated by PP2Cs thus shut down the gene expression efficiently.
| + | |
− | </p>
| + | |
− | <p>When signal 1(ON) comes, promoter RD29A drives expression of SnRK2.2, which can phosphorylate ABF2 (constitutively expressed). Comparing to protein co-facter association ,enzyme catalysis can produce a large quantity of phosphorylated ABF2 in a short time, then enhances pRD29A to turn on the gene of interest’s expression. When signal 2(OFF) comes,gene PP2CA expresses, dephosphorylating ABF2 and inactivating SnRK2.2, thus turning the system into OFF state.
| + | |
| </p> | | </p> |
| + | <img src="https://static.igem.org/mediawiki/2016/7/70/T--HUST-China--Logic-gate.jpg" class="img-responsive" alt=""> |
| + | <p>The circuit can rapidly reach its stable state because of positive feedback. And users can adjust its threshold too.</p> |
| + | <br><a href="https://2016.igem.org/Team:HUST-China/Proof#location_Pro_ver" style="text-decoration:none"><button type="button" class="btn btn-info center-block">Click to know more about the circuit</button></a> |
| + | <!-- h3二级标题 --> |
| + | <!-- <h3>SubHeading</h3> |
| + | <p>dfwef</p> |
| + | <h3>Hello world</h3> |
| + | <p>fdwfewf</p> --> |
| + | <p><strong>For more application examples:</strong></p> |
| + | <p>We provided a solution to lactose intolerance based on this toolkit.</p> |
| + | <p>The input of signals are achieved by two promoters: plac (lactose inducible) and patp2 (base inducible). As for the output, we set the gene 1 as iLDH and gene 2 as beta-galactosidase. iLDH can transform lactic acid into pyruvate whileβ-galactosidase can degrade lactose. In this way, the system can provide a promising way for the treatment of lactose intolerance.</p> |
| + | <img src="https://static.igem.org/mediawiki/2016/c/c8/T--HUST-China--description-app.png" alt=""> |
| + | <p><strong>In our bi-stable switch</strong>, users can define an open signal and off signal to control the expression of target gene. The most attractive feature of the circuit is its switch efficiency, the mechanism of which is cascade reaction. </p> |
| + | <img src="https://static.igem.org/mediawiki/2016/8/83/T--HUST-China--description-logic.png" class="img-responsive" alt=""> |
| + | <br><a href="https://2016.igem.org/Team:HUST-China/Proof#location_Euk_ver" style="text-decoration:none"><button type="button" class="btn btn-info center-block"> Click to know more about the circuit</button></a> |
| </article> | | </article> |
| </div> | | </div> |
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| <img src="https://static.igem.org/mediawiki/2016/f/f0/T--HUST-China--returnTop.png" alt=""> | | <img src="https://static.igem.org/mediawiki/2016/f/f0/T--HUST-China--returnTop.png" alt=""> |
| </div> | | </div> |
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