Team:TU Eindhoven/Project

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        <span>Introduction</span>   
        <span>Introduction</span>   
      <p class="para">This year’s iGEM team of the Eindhoven University of Technology focuses on a fundamental problem in the application of genetically modified bacteria: bacteria are not suited for many environments.
      <p class="para">This year’s iGEM team of the Eindhoven University of Technology focuses on a fundamental problem in the application of genetically modified bacteria: bacteria are not suited for many environments.
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For example: local bacterial production of medicine has a promising future as a medical treatment, but the human immune system still is a big limiting factor for this new technology. Another field that would greatly benefit from more resilient bacteria is the bioreactor industry; bacteria that can survive in high pressure and temperature environments can help increase efficiency of reactors.
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For example, local bacterial production of medicine has a promising future as a medical treatment, but the human immune system is still a big limiting factor for this new technology. Another field that would greatly benefit from more resilient bacteria is the bioreactor industry. Bacteria that can survive in high pressure and temperature environments can help increasing efficiency of reactors.
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<p class="para">To create such resilient bacteria the 2014 team has designed a ‘plug and play’ system using copper free click chemistry to attach different chemical groups to create bio-layers on E. Coli cell membranes. Circularly permuted OmpX (CPX), an outer membrane protein, was mutated to contain an azido-functionalized unnatural amino acid. CPX  functions as an anchor for any DBCO functionalized molecule to click on to. The polymers used in this project were designed to form hydrogels , which enables the bacteria to have antifouling properties.
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To create such resilient bacteria the 2014 team has designed a ‘plug and play’ system using copper free click chemistry to attach different chemical groups to create bio-layers on E. Coli cell membranes. Circularly permuted OmpX (CPX), an outer membrane protein, was mutated to contain an azido-functionalized unnatural amino acid. CPX  functions as an anchor for any DBCO functionalized molecule to click onto. The polymers used in this project were designed to form hydrogels, which enables the bacteria to have antifouling properties.
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  <span>Our Goal</span>   
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  <span>Microfluidics</span>   
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      <p class="para">Our goal is to genetically engineer an E. coli bacteria strain in which each bacterium is able to produce a hydrogel capsule around its entire cell membrane, with the help of microfluidic techniques, in order to evade the immune system. This is visualised in Figure 2. A criterion that must be met is the fact that this engineered E. coli must produce a degradable capsule, either enzymatically or after a certain induction. The advantages of this technique are a possible gelation from the inside toward the outside (a more homogeneous gelation process), a controllable cell growth (only one bacterium per capsule), and the bacterium will be able to control its own polymerisation process enzymatically – this will result in more controllable drug release. </p>
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      <p class="para">In order to precisely control the hydrogel formation, microfluidic devices are used in which the conditions are optimal to form individually encapsulated cells. This way clustering of cells is prevented and the end product will be usable beads instead of large aggregated blobs. </p>
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Latest revision as of 13:22, 12 September 2014


Project Description

Project Description

Introduction

This year’s iGEM team of the Eindhoven University of Technology focuses on a fundamental problem in the application of genetically modified bacteria: bacteria are not suited for many environments. For example, local bacterial production of medicine has a promising future as a medical treatment, but the human immune system is still a big limiting factor for this new technology. Another field that would greatly benefit from more resilient bacteria is the bioreactor industry. Bacteria that can survive in high pressure and temperature environments can help increasing efficiency of reactors.

To create such resilient bacteria the 2014 team has designed a ‘plug and play’ system using copper free click chemistry to attach different chemical groups to create bio-layers on E. Coli cell membranes. Circularly permuted OmpX (CPX), an outer membrane protein, was mutated to contain an azido-functionalized unnatural amino acid. CPX functions as an anchor for any DBCO functionalized molecule to click onto. The polymers used in this project were designed to form hydrogels, which enables the bacteria to have antifouling properties.



Application

With our team’s biomedical background in mind, an anti-fouling chemical layer for use in the human body was chosen to test the ‘plug and play’ system. An anti-fouling hydrogel has to have little to no interaction with the human immune system, thus preventing immune responses caused by the presence of bacteria. Dibenzocyclooctyne Polyethylene glycol 10kDa (DBCO-PEG 10kDa) was chosen as the molecule to click onto OmpX to form the hydrogel because it has good anti-fouling properties and the modular length allows for easy testing on a smaller scale.



Microfluidics

In order to precisely control the hydrogel formation, microfluidic devices are used in which the conditions are optimal to form individually encapsulated cells. This way clustering of cells is prevented and the end product will be usable beads instead of large aggregated blobs.

Created by iGEM Team TU Eindhoven 2014

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