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− | <p style="color: #273236"> | + | <p style="color: #273236">To express mBBI, we had to choose a biological system that was not only capable of high expression but be robust, adaptable and genetically manipulable, for ease of transport and use in remote environments. After creating a list of different types of cells which can be used for the expression of the peptide, we found that <i>Bacillus subtilis</i> is especially suited for the patch design. This is due to many reasons. Firstly <i>B. subtilis</i> is able to express high amounts of recombinant proteins making it ideal for producing and delivering the correct dosage into the user. Secondly <i>B.subtilis</i> is capable of forming spores, -which have been found in arctic permafrost dating tens of thousands of years and yet are still viable- making it easy to store in a desiccated or frozen manner. Lastly <i>B.subtilis </i>expresses high amounts of recombinase A which allows genes to be stably integrated into the chromosome. |
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+ | Our strain of B.subtilis (WB800) is particularly suited for high production because it is deficient in eight extracellular proteases which may prematurely degrade the peptide. Thus our strain will allow the peptide to be stable in the media within the patch. We conducted tests on the growth patterns of <i>B.subtilis</i> in the novel environment of a patch and optimized the patch to simulate an ideal growth environment. We have also designed a construct which provides inducible competency to the <i>B. subtilis</i> strain, making this bacterium much easier to work with for future teams. </p> | ||
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Revision as of 03:02, 18 October 2016
Our Project
Project Description
Mars and the cosmos are a tantalizingly close goal for Mankind to reach in the next century; however, there are several glaring problems that we must overcome to reach such an objective. Radiation is chief among them. On Earth, individuals annually receive 2.4 mSv of ionizing radiation (IR), which is easily tolerated by the human body. Unfortunately, without the protection of the magnetosphere, astronauts in space are exposed to high levels of IR in the range of 50 to 2000 mSv. This level of radiation causes the accumulation of deleterious double strand breaks in DNA. Despite current research into methods of IR protection, many solutions such as radiation shield coating are quite expensive, especially to transport to space. Even existing biological solutions are insufficient, such as the use of ingestible or injectable radioprotectors, which are subject to sinusuoidal pharmacokinetics. Our project is based on the administration of the naturally occuring peptide Bowman-Birk Protease Inhibitor (BBI), which has been shown to confer protection against DNA damage following radiation exposure. (Dittmann et al., 2003).
References
Dittmann, K.H., Mayer, C., and Rodemann, H.P. (2003). Radioprotection of normal tissue to improve radiotherapy: the effect of the Bowman Birk protease inhibitor. Current Medicinal Chemistry - Anticancer Agents, 3(5), 360-363.
OUR CHASSIS
To express mBBI, we had to choose a biological system that was not only capable of high expression but be robust, adaptable and genetically manipulable, for ease of transport and use in remote environments. After creating a list of different types of cells which can be used for the expression of the peptide, we found that Bacillus subtilis is especially suited for the patch design. This is due to many reasons. Firstly B. subtilis is able to express high amounts of recombinant proteins making it ideal for producing and delivering the correct dosage into the user. Secondly B.subtilis is capable of forming spores, -which have been found in arctic permafrost dating tens of thousands of years and yet are still viable- making it easy to store in a desiccated or frozen manner. Lastly B.subtilis expresses high amounts of recombinase A which allows genes to be stably integrated into the chromosome. Our strain of B.subtilis (WB800) is particularly suited for high production because it is deficient in eight extracellular proteases which may prematurely degrade the peptide. Thus our strain will allow the peptide to be stable in the media within the patch. We conducted tests on the growth patterns of B.subtilis in the novel environment of a patch and optimized the patch to simulate an ideal growth environment. We have also designed a construct which provides inducible competency to the B. subtilis strain, making this bacterium much easier to work with for future teams.
Design Overview
We designed our project to overcome common issues in existing technologies such as cost, portability and durability. Our solution is superior to current technologies because our patch is portable, lightweight, safe to use, can be stored long-term, and produces and delivers peptides at a steady rate, which solves the issues of pharmacokinetics. We worked with professionals within NASA and the Canadian Space Agency to integrate our solution with current space travel infrastructure, and with companies specializing in transdermal delivery to optimize the delivery system. In combining our biological delivery system and the professional advice we received, we are able to create a transdermal patch that can contain B. subtilis while delivering our BBI peptide.
About us
The 2016 U of C Calgary iGEM team is a multidisciplinary team based out of the University of Calgary. Our team is made up of undergraduate students from all years of study, hailing from the broad backgrounds of biology, microbiology, biomedical sciences, bioinformatics, and engineering. We are based out of the O'Brien Centre Labs within the U of C's Health Science Centre.