Team:SoundBio/Project

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Project
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Design

Project Background & Problem

Tissue engineering is the design, development, and testing of artificial tissues, typically for transplantation, that mimic native tissues enough to take their place. This field has made substantial progress in the last few decades, but " there are [still] no materials that fully capture the intricacies of the native tissue nor restore function to an ideal level ".

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Project Description & Solution

This year our project, Bacto-Basics, strives to spatially control the attachment of functional proteins to bacterial cellulose using an optogenetics, synthetic biology, and systems biology interdisciplinary study by incorporating hardware and software aspects. When selecting our primary goals we strived to stay consistent and practical, such that it would be possible to reach these goals and show our team’s progress. Under our primary umbrella project, we were split into subteams, such as public relations and wetlab, and devised goals for each subteam to achieve over the course of the iGEM 2019 season as the subteams were better informed to create their own feasible goals.

Our primary project goals this year are:
  1. Engineer E. coli to express fusion proteins, which are cellulose binding domains (CBDs) connected to chromoproteins, and control that expression via two different light sensor systems: a red light system and a blue one.
  2. Design/build a bioreactor that has minimal to no agitation so the K. Rhaeticus can grow. How we will achieve this is by testing the K. Rhaeticus in minimal to no agitating bioreactors such as wave bioreactors, static bioreactors, and airlift bioreactors.
  3. Create a model of the bioreactor processes to speed up testing and allow us to find optimal conditions for BC growth and functionalization, both in terms of the organisms and their genetic circuits, and the hardware involved.
  4. To engage our community in science by giving a mini iGEM experience to young students and incorporate public feedback and engagement with our project.

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Project Abstract

Our project aims to create a platform for precise, light-based control of bacterial cellulose (BC) functionalization for a multitude of applications including burn wound treatment, tissue scaffolding, and air filtration. We will grow Komagataeibacter rhaeticus (K. rhaeticus), a bacterial species that naturally produces BC. We will engineer E. coli to attach fusion proteins via a double cellulose binding domain for functionalization of BC. Levels of functionalization will be controlled with focused light via two optogenetic circuits utilizing red and blue light. By designing and constructing a bioreactor compatible with our optogenetic light control system, we aim to optimize K. rhaeticus growth and BC production by using Raspberry Pi-controlled sensors to monitor our culture’s pH, oxygen levels and temperature and developing a method to optimize media conditions. Our project demonstrates a proof-of-concept regarding BC functionalization through the attachment of chromoproteins to the cellulose membrane.

Future Implications

This project has the potential to increase the functionality of BC which could allow for more effective tissue scaffolding, burn treatment, and more. Adding proteins to the surface of BC can change its properties. For example, synthetic collagen is a protein that is found in the extracellular matrix of places such as bone and skin tissue. By attaching this protein to BC, BC can be given tissue scaffolding properties.

For more information on the possible BC applications please click here.

Project Design

Our team designed two genetic circuits our team designed - the red light circuit and the blue light circuit to spatially control the attachment of proteins to bacterial cellulose.
Design Design
Left: Red Light Circuit
Right: Blue Light Circuit

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Results

The FixK2 promoter was successfully characterized and experiments showed that there was little to no leaky expression in the promoter. Unfortunately, we were unable to clone the red and blue light circuits in time due to reasons such as the complexity of the circuit.

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