Difference between revisions of "Team:UNOTT/Description"

 
(103 intermediate revisions by 3 users not shown)
Line 4: Line 4:
  
 
<head>
 
<head>
 +
<meta name="viewport" content="width=device-width, initial-scale=1">
 
<meta charset="UTF-8">
 
<meta charset="UTF-8">
 
<link href="https://fonts.googleapis.com/css?family=Karla" rel="stylesheet">
 
<link href="https://fonts.googleapis.com/css?family=Karla" rel="stylesheet">
−
<style>
 
−
body {
 
−
    font-family: 'Karla', sans-serif;
 
−
}
 
  
 
<style>
 
<style>
Line 17: Line 14:
 
       ===================================*/
 
       ===================================*/
 
       body {
 
       body {
−
       color: #183532;
+
      
 
       font-family: 'Lato', sans-serif;
 
       font-family: 'Lato', sans-serif;
 
       font-weight: 400;
 
       font-weight: 400;
Line 81: Line 78:
 
       .section-two,
 
       .section-two,
 
       .section-three {
 
       .section-three {
−
      background: #fff;
+
        margin: auto;
 +
    width: 60%;
 +
    padding: 10px;
 +
    font-family: 'Karla', sans-serif;
 +
    background-color: #ffffff;
 +
     
 
       }
 
       }
  
Line 107: Line 109:
 
       .img-designers,
 
       .img-designers,
 
       .img-developers {
 
       .img-developers {
−
       height: 250px;
+
       height: 400px;
−
       width: 250px;
+
       width: 400px;
 
       background-position: center center;
 
       background-position: center center;
 
       background-size: cover;
 
       background-size: cover;
Line 158: Line 160:
 
}
 
}
  
−
      h1 {
+
   
−
          font-size: 300px;
+
−
         
+
−
    }
+
 
.maintext {
 
.maintext {
 
position: absolute;
 
position: absolute;
Line 181: Line 180:
 
.contentmargin {
 
.contentmargin {
 
     margin: auto;
 
     margin: auto;
−
     width: 65%;
+
     width: 60%;
 
     padding: 10px;
 
     padding: 10px;
 
     font-family: 'Karla', sans-serif;
 
     font-family: 'Karla', sans-serif;
 
     background-color: #ffffff;
 
     background-color: #ffffff;
 +
}
  
 +
h1 {
 +
  font-weight: normal;
 +
  letter-spacing: -1px;
 +
  color: #34495E;
 
}
 
}
 +
h2 {
 +
  font-weight: normal;
 +
  letter-spacing: -1px;
 +
  color: #34495E;
 +
}
 +
p {
 +
  font-weight: normal;
 +
  letter-spacing: -1px;
 +
  color: #4b524a;
 +
 +
.ourkey {
 +
position: relative;
 +
top:15px;
 +
}
 +
h5 {
 +
  font-weight: normal;
 +
  letter-spacing: -1px;
 +
  color: #4b524a;
 
</style>
 
</style>
 
</head>
 
</head>
Line 199: Line 221:
 
</div>
 
</div>
  
−
 
+
<br>
 +
<br>
 +
<br>
 
<!-- Section Two; -->
 
<!-- Section Two; -->
 
<section class="container section-two" id="spydesingers">
 
<section class="container section-two" id="spydesingers">
 
<div class="row">
 
<div class="row">
 
<div class="col-md-6 to-match">
 
<div class="col-md-6 to-match">
−
<h2 style="color:#4b524a;">What?</h2>
+
<h1>What?</h1>
 
<article>
 
<article>
−
<p><em>Key. coli</em> provides a new, more secure form of key for accessing content. It uses random ligations and large modular repertoires of possible components to generate unique combinations that could be the next thing in security.
+
<p><b><i>Key. coli</i> provides a new, more secure, form of key for accessing content.</b> It uses random ligations and large repertoires of possible components to generate unique combinations of expression profiles; this next generation biological key could be the next BIG thing in security; watch this space!
 +
 
 
</p>
 
</p>
−
                    <p></p>
+
                 
 
 
−
<p>
+
−
</p>
+
−
<p
+
−
</p>
+
 
</article>
 
</article>
 
</div>
 
</div>
−
<div class="col-md-6 css-img-wrapper hidden-xs hidden-sm">
+
−
<img src="https://static.igem.org/mediawiki/2017/archive/b/b0/20171026145318%21T--UNOTT--openlock.png" class="img-responsive img-circle img-designers">
+
<img class="ourkey" src="https://static.igem.org/mediawiki/2017/3/36/T--UNOTT--keyk.png" style="width:40%;height:auto;">
−
</div>
+
 
 +
 +
 
 +
 
</section>
 
</section>
 
<!--//Close Section Two -->
 
<!--//Close Section Two -->
 
 
 
 
−
<!--// Section Three -->
+
−
<section class="container section-three" id="spydevelopers">
+
<div class="contentmargin">
−
<div class="row">
+
<h1>Why?</h1>
−
<div class="col-md-6 css-img-wrapper hidden-xs hidden-sm">
+
−
<div>
+
−
<img src="https://static.igem.org/mediawiki/2017/e/e4/T--UNOTT--ourkey.png" class="img-circle img-responsive img-developers"></div>
+
<p><b>Major hacking incidents are increasingly common,</b> with accounts being hacked and sensitive information stolen. Many companies are moving away from conventional passwords, which are proving to be unreliable in the hands of the public. Banks are now using physical biometric authentication procedures to correctly identify account owners. This new direction opens a market for biological “passwords”. An ideal system would be as separate from online software programs as possible while maintaining the complexity and uniqueness of a biometric system. Cells are effectively living computers, so we can programme cells to act as a changeable biometric password. </p>
−
</div>
+
−
<div class="col-md-6">
+
−
<h2 style="color:#4b524a;">Current issues with security</h2>
+
−
<article>
+
−
<p>
+
−
Major hacking incidents are increasingly common, with accounts being hacked and sensitive information stolen. Many companies are moving away from conventional passwords, which are proving to be unreliable in the hands of the public. Banks are now using physical biometric authentication procedures to correctly identify account owners. This new direction opens a market for biological “passwords”. An ideal system would be as separate from online software programs as possible, while maintaining the complexity and uniqueness of a biometric system. Cells are effectively living computers so we can programme cells to act as a changeable biometric password.
+
−
</p>
+
−
<p>
+
−
</p>
+
−
<p>
+
−
</p>
+
−
</article>
+
−
</div>
+
−
</div>
+
−
</section>
+
−
<!--//Close Section Three -->
+
  
  
−
 
−
 
−
<div class="contentmargin">
 
  
−
<h2 style="color:#4b524a;">Fluorescent proteins</h2>
+
 
 +
 
 +
<h1>How?</h1>
 +
 
 +
<p><b>A key must be unique, measurable and unpredictable.</b> In <i>Key. coli</i>, all these requirements are achieved by the random generation of modular vectors that are expressed in Escherichia coli to produce a unique and detectable fluorescent pattern. This pattern is obtained when different fluorescent proteins (GFP, RFP, CFP) and various promoters, subjected to transcription interference by dCas9, are randomly combined during ligation and transformed into the cells to generate the key. </p>
 +
<p><span style="color: #ffffff;">&nbsp;</span></p>
 +
<p><span style="color: #ffffff;">&nbsp;</span></p>
 +
 
 +
<img src="https://static.igem.org/mediawiki/2017/8/84/UNOTT2017-How1.png" alt="" width="100%" height="100%">
 +
<h5><b> Figure 1:</b> Two-plasmid modular process used to generate random<i>Key. coli</i> construct(s)<p>
 +
<p><span style="color: #ffffff;">&nbsp;</span></p>
 +
<p><span style="color: #ffffff;">&nbsp;</span></p><p><span style="color: #ffffff;">&nbsp;</span></p>
 +
<p><span style="color: #ffffff;">&nbsp;</span></p>
 +
<img src="https://static.igem.org/mediawiki/2017/b/bc/UNOTT2017-How2.png" alt="" width="100%" height="100%"></h5>
 +
 
 +
<h5><b>Figure 2:</b> Random ligation process and colony picking allows large numbers of plasmid variants to be created for use in keys. </p> <p><span style="color: #ffffff;">&nbsp;</span></p></h5>
 +
<p><span style="color: #ffffff;">&nbsp;</span></p>
 +
<br>
 +
<p>A key transport device, based on freeze-dried <i>Key. coli</i>, allows the bacteria to survive and be transported anywhere with ease. Once entry to a lock is desired, the <i>Key. coli</i> device can be activated, and the output read in a suitable detection device. </p>
 +
 
 +
<h1><i>Key. coli</i> Summary</h1>
 
 
−
<p>Fluorescent proteins are routinely used in molecular biology, are well understood and safe to use. We used 3 different fluorescent proteins (FPs) to generate unique spectra as their detection is relatively simple compared to other proteins. The potential colours of fluorescent proteins are finite. We chose to extend our potential combinations by introducing variation in three ways, as follows.</p>
 
−
                 
 
−
 
−
 
−
 
−
 
−
<h2 style="color:#4b524a;">Promoter strength</h2>
 
−
 
−
<p> We have taken 5 promoters from literature<sup>1</sup> and attached them to these fluorescent proteins. From our results you can see that they are different strengths themselves. In the future, we could have 100s of promoters with different strengths to give different fluorescence intensity.</p>
 
 
 
−
+
<p><img src="https://static.igem.org/mediawiki/2017/8/83/UNOTT2017-summary.png" alt="" width="100%" height="auto" /></p>
−
<h2 style="color:#4b524a;">Expanding the repertoire of colours and proteins</h2>
+
<br>
−
+
<br>
−
<p>Our initial proof of concept uses 3 FPs. For future expansion, there is a vast repertoire of fluorescent proteins but also any other protein could be substituted in and measured using other methods, for example by mass spec. Our assembly method is all interchangeable so any protein can be linked to any promoter and placed in any position in the plasmid. In this way, this system could be used for a vast range of applications.</p>
+
<br>
−
                   
+
<br>
−
+
−
<h2 style="color:#4b524a;">CRISPRi</h2>
+
−
+
−
<p>dCas9 is an mutated version of the popular gene editing enzyme, Cas9, that has become extremely popular since its first use in the early 2010s. It will be targeted to repress a given promoter preceding a protein by using a short guide RNA (sgRNA) which has a seed region complementary to this promoter. We have taken five promoter-sgRNA pairings from literature1 to give ON/OFF switches for fluorescent proteins. Our results show that these gRNAs have differing repression efficiencies as they are, but other work has shown that single nucleotide changes in the seed region can alter the gRNAs repression efficiency.</p>
+
 
 
−
 
−
 
−
<h2 style="color:#4b524a;">What?</h2>
 
−
<p><em>Therefore, not only can we create a plasmid that can contain any measurable protein, but we can randomly assign a promoter strength, and use CRISPRi to further modify these levels from a separate plasmid.</em>
 
−
 
−
 
−
 
−
 
−
<h2 style="color:#4b524a;">Measurement & Modelling</h2>
 
−
 
−
<p> A lock would use a fluorescence detection device that would require the keyholder to have an exact copy of the reference bacteria that the detection device uses, as the fluorescence of both samples would be compared for likeness. This is where the key transport device came in as it needed to be compact and portable, yet allow the bacteria to survive and be read easily</p>
 
−
 
−
<p>Also, in order to have reproducibility and allow accurate authentication by the keyholder, the two signals need to have minimal variance. If not, then the accuracy and completeness of emission signals is invalidated and fewer discernible combinations are possible. Therefore, we are using the data collected to form models to predict the outcomes of fluorescence under all possible conditions and allow each key to be categorized separately by their output.</p>
 
−
<
 
  
 
</div>
 
</div>

Latest revision as of 03:45, 2 November 2017

 

 

 

PROJECT DESCRIPTION

 




What?

Key. coli provides a new, more secure, form of key for accessing content. It uses random ligations and large repertoires of possible components to generate unique combinations of expression profiles; this next generation biological key could be the next BIG thing in security; watch this space!

Why?

Major hacking incidents are increasingly common, with accounts being hacked and sensitive information stolen. Many companies are moving away from conventional passwords, which are proving to be unreliable in the hands of the public. Banks are now using physical biometric authentication procedures to correctly identify account owners. This new direction opens a market for biological “passwords”. An ideal system would be as separate from online software programs as possible while maintaining the complexity and uniqueness of a biometric system. Cells are effectively living computers, so we can programme cells to act as a changeable biometric password.

How?

A key must be unique, measurable and unpredictable. In Key. coli, all these requirements are achieved by the random generation of modular vectors that are expressed in Escherichia coli to produce a unique and detectable fluorescent pattern. This pattern is obtained when different fluorescent proteins (GFP, RFP, CFP) and various promoters, subjected to transcription interference by dCas9, are randomly combined during ligation and transformed into the cells to generate the key.

 

 

Figure 1: Two-plasmid modular process used to generate randomKey. coli construct(s)

 

 

 

 

Figure 2: Random ligation process and colony picking allows large numbers of plasmid variants to be created for use in keys.

 

 


A key transport device, based on freeze-dried Key. coli, allows the bacteria to survive and be transported anywhere with ease. Once entry to a lock is desired, the Key. coli device can be activated, and the output read in a suitable detection device.

Key. coli Summary