Difference between revisions of "Team:UNOTT/Description"

 
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<h1>Description</h1>
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<p>Due to the recent explosion in hacking incidents, we were interested at looking into how biological systems can be used to overcome the limitations of current passwords. Although there was the possibility to use DNA as a password, the speed of current sequencing technologies and lack of options for creating randomness without mutating the bacteria to a point where it could not survive were limiting to the success of this option. For that reason we turned to using physical properties of bacteria such as its metabolome in order to create a biological password.</p>
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<h5>The idea</h5>
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<li> The first biological password that changes over time!
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We are looking into transforming bacteria with a unique array of existing iGEM systems to produce a unique signal of secondary metabolites, initially using fluorescence as a proof of concept. Eventually, we will use the system to produce a unique and random configuration of secondary metabolites, as our "key". In order to produce this randomness, alteration of the activity or presence of promoters associated with these metabolites will be applied using one of a few methods currently being considered by the team.</li>
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<li>This key will be used to open locked mechanism such as safes and secure doors. We see a system where the measurement of key engineered metabolites such as volatiles will give a distinct mass spectrum. A juxtaposition of a detection technique such as gas chromotography-mass spectrometry and data comparison software will compare the secondary metabolites of the "key" bacteria to the "reference/lock" from which it was taken. If the spectra of both colonies exceeds a threshold of similarity the locked object will become unlocked.</li>
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<li>After a certain amount of time, our Key will have to be renewed from the Lock colony, and when this occurs the configuration of the key is shuffled once again to ensure the key and lock are changing. </li>
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<h5>Bacterial Key Transport</h5>
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<p>There is a need for a transport mechanism for the key. This presents problems depending on the bacteria used.</p>
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<p> In Ecoli Our key transport system would need to: </p>
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<li>Keep our colonies alive for a few days. </li>
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<li>Potentially could freeze. Freezing is one of the best ways to store bacteria. </li>
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<li>The lower the temperature the longer the culture will retain viable cells.</li>
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<li>PROBLEM: Ice can damage cells due to localised accumulation of salt, it can also rupture membranes. </li>
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<li>SOLUTION: Use glycerol as a cryoprotectant </li>
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<p>For the key to be practical it would need to be portable, this is where the design of our key transport device comes in. We are currently looking into a system of a similar design to a chemostat where a continual supply of medium will allow maintenance of a culture. Other options we are looking into include the use of freeze-dried cells or microfluidics.</p>
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<h5> Project Overview </h5>
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<p>FOR CHRIS: https://www.youtube.com/watch?v=otCpCn0l4Wo</p>
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<h5> Creation of distinct spectra of different metabolite levels </h5>
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<h5> Promoter selection </h5>
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<p>We would select a range of possible promoters to give a wide variety of product expression levels.</p>
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<p>We are looking at the following methods to achieve a random selection of product levels within any given bacteria:</p>
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<li>1. Transposon shuffling of promoters between products. </li>
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<li>2. dCas9 and a randomly selected gRNA from a library of gRNAs that can interfere differentially with the promoters associated with products.</li>
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<h5> Transposons </h5>
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<p>Tn7 Transposon used due to specific target site selection for its transposition, which is impossible in other transposon species, without this modular increases in promoter activity could not be achieved as random insertions would create a gradient rather than step wise expression pattern of proteins.</p>
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<h5>dCas9 and gRNA library</h5>
 
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<h2>Possible Metabolites</h2>
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<h4>The metabolites that will be coded for by our constructs will be those previously registered with iGEM by previous teams,
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as the focus of this project falls on the assortment and variety of expression levels rather than the specific product. Nevertheless, we are looking into selecting various products that will be detectable by techniques such as GCMS and will not interact with eachother. We will also choose products that are easily distinguishable from eachother on spectra. Currently our list of potential metabolites includes the following:</h4>
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<li> product x by team x (insert wiki link here)</li>
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<p><span style="color: #ffffff;">&nbsp;</span></p>
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<p><span style="color: #ffffff;">&nbsp;</span></p>
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<p><span style="color: #ffffff;">&nbsp;</span></p>
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<h1 style="text-align: center;"><span style="color: #ffffff;">PROJECT DESCRIPTION</span></h1>
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<p><span style="color: #ffffff;">&nbsp;</span></p>
 
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<h1>What?</h1>
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<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!
  
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<img class="ourkey" src="https://static.igem.org/mediawiki/2017/3/36/T--UNOTT--keyk.png" style="width:40%;height:auto;">
  
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<h1>Why?</h1>
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<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>
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<h1>How?</h1>
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<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>
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<img src="https://static.igem.org/mediawiki/2017/8/84/UNOTT2017-How1.png" alt="" width="100%" height="100%">
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<h5><b> Figure 1:</b> Two-plasmid modular process used to generate random<i>Key. coli</i> construct(s)<p>
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<p><span style="color: #ffffff;">&nbsp;</span></p>
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<p><span style="color: #ffffff;">&nbsp;</span></p><p><span style="color: #ffffff;">&nbsp;</span></p>
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<img src="https://static.igem.org/mediawiki/2017/b/bc/UNOTT2017-How2.png" alt="" width="100%" height="100%"></h5>
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<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>
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<p><span style="color: #ffffff;">&nbsp;</span></p>
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<br>
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<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>
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<h1><i>Key. coli</i> Summary</h1>
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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