Team:Caltech/Project

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Background and Motivation

One of synthetic biology's chief promises is the potential to construct artificial, self-regulatory genetic circuits. While intracellular genetic regulation is very well-documented, signalling between cells, mediating intercellular regulation, remains less well-known and explored. This year, the Caltech 2014 iGEM team focused on the implementation of non-endogenous bacterial quorum-sensing systems in E. coli, in hopes that successful transfer of these systems into E. coli will promote enhanced prototyping of intercellular—in addition to intracellular—signaling and regulation in synthetic biology’s favorite model organism.

The original motivation for this project was to create a model system that will resemble the hormone regulatory systems of the body. Examples of disorders of regulation can be seen in diabetes, hyperthyroidism, Cushing’s disease, etc. The original intent of the project was to create an analogous model of one of these regulatory systems in E. coli, showing one plausible framework to serve as a scaffold for further research.

Project Design

The Caltech iGEM team plans to use the principles and methods of synthetic biology to create a model system that will follow the design in the picture below. The gene circuit has been designed so that "Cell 1" will produce a signaling peptide upon induction with aTc. This signaling peptide will then attach to the receptor of "Cell 2," triggering a very simple signal transduction pathway leading to expression of some target gene that, for our testing purposes, will be the reporter protein GFP. The quorum sensing systems being imported into the E. coli cells come from Gram-positive species of bacteria, which minimizes the possibility that their activity will interfere with or be interfered with by E. coli's endogenous signaling and reception systems but also introduces the challenge of trying to implement the relevant membrane proteins in an environment that significantly differs from their native environments (owing to the differences in membrane composition between Gram-positive and Gram-negative bacteria).

Our project, however, is not without precedent: we will be building off of [insert team name] team’s project in [year], which managed successfully to implement the signal reception components of the agrBCDA quorum sensing system in E. coli.

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 One of synthetic biology's chief promises is the potential to construct artificial, self-regulatory genetic circuits. While intracellular genetic regulation is very well-documented, signalling between cells, mediating intercellular regulation, remains less well-known and explored. This year, the Caltech 2014 iGEM team focused on the implementation of non-endogenous bacterial quorum-sensing systems in E. coli, in hopes that successful transfer of these systems into E. coli will promote enhanced prototyping of intercellular—in addition to intracellular—signaling and regulation in synthetic biology’s favorite model organism.
 <p> The original motivation for this project was to create a model system that will resemble the hormone regulatory systems of the body. Examples of disorders of regulation can be seen in diabetes, hyperthyroidism, Cushing’s disease, etc. The original intent of the project was to create an analogous model of one of these regulatory systems in E. coli, showing one plausible framework to serve as a scaffold for further research.</p>
-<p> The Caltech iGEM team plans to use the principles and methods of synthetic biology to create a model system  The model system will follow the design in the picture below. The gene circuit has been designed to produce a signaling peptide in response to some factor, and then have another cell respond to the produced signaling peptide to form a regulatory network. The pathway used to activate the regulation will ideally not interfere with any of the intercellular genetic circuitry already present in E. coli. The synthetic pathway will be assembled from systems that are present in other bacteria and will hopefully be dissimilar enough to not interact with any already present E. coli circuitry. We will be building off of a previous iGEM team’s project of bacterial quorum sensing and regulation in E. coli. </p>
+<h3>Project Design</h3>
+<p> The Caltech iGEM team plans to use the principles and methods of synthetic biology to create a model system that will follow the design in the picture below. The gene circuit has been designed so that "Cell 1" will produce a signaling peptide upon induction with aTc. This signaling peptide will then attach to the receptor of "Cell 2," triggering a very simple signal transduction pathway leading to expression of some target gene that, for our testing purposes, will be the reporter protein GFP. The quorum sensing systems being imported into the <i>E. coli</i> cells come from Gram-positive species of bacteria, which minimizes the possibility that their activity will interfere with or be interfered with by <i>E. coli's</i> endogenous signaling and reception systems but also introduces the challenge of trying to implement the relevant membrane proteins in an environment that significantly differs from their native environments (owing to the differences in membrane composition between Gram-positive and Gram-negative bacteria).</p>
+Our project, however, is not without precedent: we will be building off of [insert team name] team’s project in [year], which managed successfully to implement the signal reception components of the agrBCDA quorum sensing system in <i>E. coli</i>. </p>
   <img src= "https://static.igem.org/mediawiki/2014/1/11/New_system.png" width=100%> </center>
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