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| | <link rel="stylesheet" type="text/css" href="https://2015.igem.org/Template:Peking/Blue/new?action=raw&ctype=text/css " title="blue" media="screen" /> | | <link rel="stylesheet" type="text/css" href="https://2015.igem.org/Template:Peking/Blue/new?action=raw&ctype=text/css " title="blue" media="screen" /> |
| | <style type="text/css">/*get rid of the annoying iGEM style*/ | | <style type="text/css">/*get rid of the annoying iGEM style*/ |
| | + | #top_menu_inside, #top_menu_inside:before, #top_menu_inside:after { -webkit-box-sizing: content-box; -moz-box-sizing: border-box; box-sizing: content-box;} |
| | + | #top_menu_under{font-size: 12px;} |
| | + | #top_menu_14{font-size: 12px; height:18px} |
| | + | #top_menu_inside li{font-size: 12px;} |
| | + | #user_item{font-size: 12px;} |
| | #content{padding:0;border:0;width:100%;} | | #content{padding:0;border:0;width:100%;} |
| | .firstHeading{visibility:hidden;} | | .firstHeading{visibility:hidden;} |
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| | background-color:#e0e0e0;margin-top:1%;margin-bottom:20px;padding:20px;padding-top:40px; | | background-color:#e0e0e0;margin-top:1%;margin-bottom:20px;padding:20px;padding-top:40px; |
| | } | | } |
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| | </style> | | </style> |
| | </head> | | </head> |
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| | <div class="navbar-collapse collapse" style="padding-top:5px;padding-bottom:0"> | | <div class="navbar-collapse collapse" style="padding-top:5px;padding-bottom:0"> |
| | <!-- Start Navigation List --> | | <!-- Start Navigation List --> |
| − | <ul class="nav navbar-nav navbar-right " style="padding-bottom:15px;height:88px;padding-top:10px"> | + | <ul class="nav navbar-nav navbar-right " style="padding-bottom:15px;padding-top:10px"> |
| | <li> | | <li> |
| | <a href="https://2015.igem.org/Team:Peking/JudgingCriteria">Achievements</a> | | <a href="https://2015.igem.org/Team:Peking/JudgingCriteria">Achievements</a> |
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| | <div class="row"> | | <div class="row"> |
| | <div class="col-md-6"> | | <div class="col-md-6"> |
| − | <h2 style="font-size:20px; margin-bottom:5px; padding-bottom:0">Project</h2> | + | <h2 style="font-size:20px; margin-bottom:5px; padding-bottom:0">P<span style="text-transform:lowercase">roject</span></h2> |
| | <p style="margin-top:0px;font-size:14px">It takes half your life before you discover life is a do-it-yourself project.</p> | | <p style="margin-top:0px;font-size:14px">It takes half your life before you discover life is a do-it-yourself project.</p> |
| | </div> | | </div> |
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| | <!--End sidebar--> | | <!--End sidebar--> |
| | <!-- Page Content --> | | <!-- Page Content --> |
| − | <div class="col-md-9 page-content"> | + | <div class="col-md-9 page-content" style="text-align:justify;"> |
| | <div id="project"> | | <div id="project"> |
| | <!-- Classic Heading --> | | <!-- Classic Heading --> |
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| | <h3 class="classic-title" style="margin-top:50px"><span><a style="color:#00afd1;" href="#">Overview</a></span></h3> | | <h3 class="classic-title" style="margin-top:50px"><span><a style="color:#00afd1;" href="#">Overview</a></span></h3> |
| | <div style="margin-top:30px; margin-bottom:0"> | | <div style="margin-top:30px; margin-bottom:0"> |
| − | <p>Tuberculosis (TB), caused by Mycobacterium tuberculosis, remains one of the world's most serious public health problems. Although tuberculosis is curable and the treatment success rate is high, it is still the second most common cause of death from infectious disease. Most of the deaths occur for lack of effective identification of those in need of therapy. Case detection is currently the rate-limiting step in TB control.</p> | + | <p>Tuberculosis (TB), caused by <I>Mycobacterium tuberculosis</I>, remains one of the world's most serious public health problems. Although tuberculosis is curable and the treatment success rate is high, it is still the second most common cause of death from infectious disease. Most of the deaths occur for the lack of effective identification of those in need of therapy. Case detection is currently the rate-limiting step in TB control.</p> |
| − | <p>The currently major TB detection methods all have their own problems. Nucleic Acid Detection (NAD) is a safer, faster, and more sensitive detection method available. However, it has short comes: the high false positive rate from non-specific amplification, and the requirement of extremely expensive clumsy instruments, which make NAD not common for TB diagnosis. </p> | + | <p>The currently widely-used TB detection methods all have their own problems. Nucleic Acid Detection (NAD) is a safer, faster, and more sensitive detection method. However, its shortcoming is critical: the high false-positive rate from non-specific amplification, and the requirement of extremely expensive clumsy instruments; these make NAD not common for TB diagnosis. </p> |
| − | <p>To obviate such problems, Peking iGEM developed a novel detection system that can directly read out specific sequence information inside the PCR amplified product, and convert the information into visible signal. Combined with our work in multi-marker array design and hardware development, this new advanced system can be turned out as a powerful tool in TB diagnosis, with a huge potential for applications and extensions.</p> | + | <p>To obviate such shortcoming, Peking iGEM developed a novel detection system, paired dCas9 (PC) reporter, that converts the sequence-specific information of pathogenic bacteria's genome (in our case, <i>M. tuberculosis</i>, MTB) into easily readable signal including bioluminescence, pigment, or electric current. Our PC Reporter was successfully applied to the detection of <span style="color:#00afd1;"><B>real pathogenic <I>M.Tuberculosis</I> H37Rv</B></span> (<b>isolated genomic DNA that is absolutely safe, prepared by our collaborator, not by us</b>). Combined with our work in multiple-marker array (using MTB-specific markers extracted from the entire genome of MTB) and hardware development, our PC reporter system is expected to be a powerful tool in TB diagnosis, with a huge potential for various applications and extensions.</p> |
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| | </div> | | </div> |
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| − | <h3 class="classic-title" style="margin-top:50px"><span><a style="color:#00afd1;" href="https://2015.igem.org/Team:Peking/Design/PC_Reporter">PC Reporter</a></span></h3> | + | <h3 class="classic-title" style="margin-top:50px"><span><a style="color:#00afd1;" href="https://2015.igem.org/Team:Peking/Design/PC_Reporter">Paired dCas9 reporter</a></span></h3> |
| | <div style="margin-top:30px; margin-bottom:0"> | | <div style="margin-top:30px; margin-bottom:0"> |
| | <div class="row"> | | <div class="row"> |
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| | </div> | | </div> |
| | <div class="col-md-6"> | | <div class="col-md-6"> |
| − | <p> With two fragments of split firefly luciferase separately fused to dCas9 protein, we successfully designed and built a paired dCas9 reporter (PC reporter) that can convert specific sequence information into light signal. We finished the characterization and optimization. Most importantly, we successfully demonstrated the PC reporter with isolated genomic DNA from a pathogenic MTB.</p> | + | <p> It is well known that CRISPR/dCas9 has a unique ability to be programmed to bind any sequence with the assistance of sgRNA; it was conventionally used for DNA editing or genome study. In our project, however, we integrate split reporters into CRISPR/Cas9 by translationally fusing two fragments of a split reporter to dCas9, respectively, to convert the sequence-specific information of pathogenic bacteria's genome (in our case, <i>M. tuberculosis</i>) into easily readable signal including bioluminescence or pigment. We demonstrated that the PC reporter is highly compatible with NAD-based diagnosis using isolated genomic DNA of MTB.</p> |
| | </div> | | </div> |
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| | </div> | | </div> |
| | <div class="col-md-6"> | | <div class="col-md-6"> |
| − | <p>To deal with challenges from clinical practices, such as strain mutations, sample variations, and other uncontrollable environmental factors, we designed an array to extract more sequence information for our PC Reporter to test multiple sites on the target genome at one time to improve the reliability. With statistics analysis, we can analyze read-outs in a quantitative way.</p> | + | <p>To deal with challenges from clinical practices, such as strain mutations, sample variations, and other uncontrollable environmental factors, we designed an array to extract sequence information from the entire genome of <i>M. tuberculosis</i>, for our PC Reporter to measure multiple sites on the target genome at one time, thus to improve the reliability of diagnosis. By statistics analysis, we are able to present the readouts in a quantitative way.</p> |
| | </div> | | </div> |
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| | </div> | | </div> |
| | <div class="col-md-6"> | | <div class="col-md-6"> |
| − | <p>Noticing that most TB cases occur in developing areas, we designed and built a device (prototype), which was portable, affordable, and can be easily used by medical workers.</p> | + | <p>Noticing that most TB cases occur in developing areas, we built an electronic device (despite prototype), which was portable, affordable, and easy to use for local medical workers.</p> |
| | </div> | | </div> |
| | </div> | | </div> |
| | </div> | | </div> |
| − | <h3 class="classic-title" style="margin-top:50px"><span><a style="color:#00afd1;" href=""https://2015.igem.org/Team:Peking/Design/Isothermal>Specific isothermal amplification</a></span></h3> | + | <h3 class="classic-title" style="margin-top:50px"><span><a style="color:#00afd1;" href=""https://2015.igem.org/Team:Peking/Design/Isothermal>Simplified isothermal amplification</a></span></h3> |
| | <div style="margin-top:30px; margin-bottom:0"> | | <div style="margin-top:30px; margin-bottom:0"> |
| | <div class="row"> | | <div class="row"> |
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| | </div> | | </div> |
| | <div class="col-md-6"> | | <div class="col-md-6"> |
| − | <p>In order to popularize our detection system to developing areas, we invented two novel isothermal amplification methods that can use one pair of easily designed primers to achieve specific amplification.</p> | + | <p>In order to popularize our diagnosis method in developing areas, we invented two novel isothermal amplification methods whose mechanism and operation requirements have been significantly simplified.</p> |
| | </div> | | </div> |
| | </div> | | </div> |
