Team:UC Chile/Cyanolux/Project short

From 2012.igem.org

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{{UC_Chile4}}
{{UC_Chile4}}
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Main Goal:
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<h1>Main Goal:</h1>
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Our project consists on achieving bioluminescence controlled under circadian rhythms with long-term functionality. To do this, we have thought on coupling the bioluminescence pathway with endogenous circadian rhythms only during dusk hours, so that during daytime there is no production of light. Our aim is to produce a bioluminescent cyanobacteria which lights up during dusk hours and that regenerates the substrates during the day.
Our project consists on achieving bioluminescence controlled under circadian rhythms with long-term functionality. To do this, we have thought on coupling the bioluminescence pathway with endogenous circadian rhythms only during dusk hours, so that during daytime there is no production of light. Our aim is to produce a bioluminescent cyanobacteria which lights up during dusk hours and that regenerates the substrates during the day.
We believe that our project will serve as a proof of concept about developing high-level functionality from biology in the form of a biological lamp, Luxilla biolamp, which turns on only during the night and recharges itself during the day.
We believe that our project will serve as a proof of concept about developing high-level functionality from biology in the form of a biological lamp, Luxilla biolamp, which turns on only during the night and recharges itself during the day.
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Rationale:
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<h2>Rationale:</h2>
The importance of Biological context in Synthetic Biology has been largely underestimated. We have addressed this issue by centering our project on enhancing functionality of a previoulsy characterized Biobrick, LuxBrick, by placing it in a context which allows new features.
The importance of Biological context in Synthetic Biology has been largely underestimated. We have addressed this issue by centering our project on enhancing functionality of a previoulsy characterized Biobrick, LuxBrick, by placing it in a context which allows new features.
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Bioluminescence
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<h3>Bioluminescence</h3>
Bioluminescence is a enzymatic reaction which yield photons in the process, producing light.  
Bioluminescence is a enzymatic reaction which yield photons in the process, producing light.  
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In 2010 the Cambridge iGEM team Biobricked the LuxBrick, a collection of genes from the Lux operon that incorporate both the Luciferase and the substrate production enzymes, allowing endogenous bioluminescence on E. coli
In 2010 the Cambridge iGEM team Biobricked the LuxBrick, a collection of genes from the Lux operon that incorporate both the Luciferase and the substrate production enzymes, allowing endogenous bioluminescence on E. coli
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Chassis
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<h3>Chassis</h3>
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We have chosen to work with a model cyanobacteria, Synechocystis PCC. 6803 because it exhibits autotrophic metabolism and circadian rhythms.  
We have chosen to work with a model cyanobacteria, Synechocystis PCC. 6803 because it exhibits autotrophic metabolism and circadian rhythms.  
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Coupling the endogenous circadian rhythms of this organism to the expression of the Lux genes will enable high-level functionality through a complex behaviour.  
Coupling the endogenous circadian rhythms of this organism to the expression of the Lux genes will enable high-level functionality through a complex behaviour.  
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<h2>Strategy</h2>
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Strategy
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According to literature (CITA!), the liminting step for the bacterial bioluminescent reaction is the substrate (n-decanal) concentration, therefore, to control the light emition over time we decided to control it´s abundance in the cells, which in our model is a function of the substrates generation (by Lux C, D, E and G enzymes) and consumption (by the LuxAB luciferase).
According to literature (CITA!), the liminting step for the bacterial bioluminescent reaction is the substrate (n-decanal) concentration, therefore, to control the light emition over time we decided to control it´s abundance in the cells, which in our model is a function of the substrates generation (by Lux C, D, E and G enzymes) and consumption (by the LuxAB luciferase).
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In turn, the production of these enzymes can be specifically set to any desired  time of the day by fusing their CDSs to promoters controlled by the cyrcadian rythm.
In turn, the production of these enzymes can be specifically set to any desired  time of the day by fusing their CDSs to promoters controlled by the cyrcadian rythm.
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Mathematical Modelling.
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<h3>Mathematical Modelling</h3>
Our model works as a “black box” in which the input takes the form of a specific hour of the day (i.e the hour on which you want your metabolite to reach maximal concentration) and the output is a couple of promoters from Synechocystis genome.  
Our model works as a “black box” in which the input takes the form of a specific hour of the day (i.e the hour on which you want your metabolite to reach maximal concentration) and the output is a couple of promoters from Synechocystis genome.  
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for more details please check (link: mathematical model)
for more details please check (link: mathematical model)
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Wetlab strategy
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<h3>Wetlab strategy</h3>
Having chosen the right promoters we set out to built our constructs to transform synechocystis.
Having chosen the right promoters we set out to built our constructs to transform synechocystis.
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-Imagen?-
-Imagen?-
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Implementation
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<h2>Implementation</h2>
Synthetic biology inspires in nature to make abstractions of it´s principles and mechanisms.v
Synthetic biology inspires in nature to make abstractions of it´s principles and mechanisms.v

Revision as of 22:14, 25 October 2012

Project: Luxilla - Pontificia Universidad Católica de Chile, iGEM 2012