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<p class="c3 c11"><span style="overflow: hidden; display: inline-block; margin: 0.00px 0.00px; border: 0.00px solid #000000; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px); width: 418.67px; height: 217.33px;"><img alt="" src="https://static.igem.org/mediawiki/2016/4/4a/T--UofC_Calgary--Syed00.jpg" style="width: 418.67px; height: 217.33px; margin-left: 0.00px; margin-top: 0.00px; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px);" title=""></span> | <p class="c3 c11"><span style="overflow: hidden; display: inline-block; margin: 0.00px 0.00px; border: 0.00px solid #000000; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px); width: 418.67px; height: 217.33px;"><img alt="" src="https://static.igem.org/mediawiki/2016/4/4a/T--UofC_Calgary--Syed00.jpg" style="width: 418.67px; height: 217.33px; margin-left: 0.00px; margin-top: 0.00px; transform: rotate(0.00rad) translateZ(0px); -webkit-transform: rotate(0.00rad) translateZ(0px);" title=""></span> | ||
</p> | </p> | ||
− | <h3 | + | <h3><span>Patch Design</span></h3> |
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<li class="c0"><span>The circular design of the packet containing media and spores, encourages equal distribution of liquid pressure around the circumference. Since the packets are made up of flexible polyethylene material, this design is more advantageous when cases such as accidental bumping or ‘squishing’ of the patch happen by distributing the force.</span> | <li class="c0"><span>The circular design of the packet containing media and spores, encourages equal distribution of liquid pressure around the circumference. Since the packets are made up of flexible polyethylene material, this design is more advantageous when cases such as accidental bumping or ‘squishing’ of the patch happen by distributing the force.</span> | ||
</li> | </li> | ||
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<li class="c0"><span>The fact that our patch is also non-invasive and only present in the external part of the body promotes safety better than an implant.</span> | <li class="c0"><span>The fact that our patch is also non-invasive and only present in the external part of the body promotes safety better than an implant.</span> | ||
</li> | </li> | ||
− | <h3 | + | <h3><span>Choosing Patch Materials</span></h3> |
<li class="c0"><span>All materials used in the patch is chemically compatible and biocompatible, which poses less risks of user immune response.</span> | <li class="c0"><span>All materials used in the patch is chemically compatible and biocompatible, which poses less risks of user immune response.</span> | ||
</li> | </li> | ||
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</li> | </li> | ||
− | <h3 | + | <h3><span >Containment</span></h3> |
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<li class="c0"><span class="c1">Contamination issues may be a problem in space if the bacteria were to escape. To protect against the event that our radioprotective patch causes contamination in space or on earth, we would engineer inducible kill switches that could eradicate the bacteria if need be. Additionally, engineering controls such as the protective physical barriers of the patch would prevent the outbreak of bacteria. Various safety mechanisms have been included in the device and they are listed below.</span> | <li class="c0"><span class="c1">Contamination issues may be a problem in space if the bacteria were to escape. To protect against the event that our radioprotective patch causes contamination in space or on earth, we would engineer inducible kill switches that could eradicate the bacteria if need be. Additionally, engineering controls such as the protective physical barriers of the patch would prevent the outbreak of bacteria. Various safety mechanisms have been included in the device and they are listed below.</span> | ||
</li> | </li> | ||
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<li class="c0"><span class="c1">Public safety: Auxotrophy using Threonine was integrated in </span><span class="c1 c8">Bacillus</span><span class="c1"> making it unlikely it will survive outside of lab.</span> | <li class="c0"><span class="c1">Public safety: Auxotrophy using Threonine was integrated in </span><span class="c1 c8">Bacillus</span><span class="c1"> making it unlikely it will survive outside of lab.</span> | ||
</li> | </li> | ||
<li class="c0"><span class="c1">Environmental safety: It’s possible our </span><span class="c8 c1">Bacillus</span><span class="c1"> incorporated with BBI could possibly outcompete native microbes, however the the bacteria’s survivability outside the lab has not been increased enough to make a significant difference</span> | <li class="c0"><span class="c1">Environmental safety: It’s possible our </span><span class="c8 c1">Bacillus</span><span class="c1"> incorporated with BBI could possibly outcompete native microbes, however the the bacteria’s survivability outside the lab has not been increased enough to make a significant difference</span> | ||
</li> | </li> | ||
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<li class="c0"><span>The backing layer and the size-controlling membrane prevents the bacteria from escaping outside of the patch and being released onto the skin, respectively.</span> | <li class="c0"><span>The backing layer and the size-controlling membrane prevents the bacteria from escaping outside of the patch and being released onto the skin, respectively.</span> | ||
</li> | </li> | ||
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<li class="c0"><span>See pages 15 - 16 of the Device Manual to tackle contamination issues.</span> | <li class="c0"><span>See pages 15 - 16 of the Device Manual to tackle contamination issues.</span> | ||
</li> | </li> | ||
− | |||
<li class="c0"><span class="c1">The engineered organism could potentially disrupt existing ecologies and be difficult to contain; however, as mentioned previously, integrated BBI would not make a difference in the bacteria’s survivability outside the lab, and we have taken care to ensure that the Threonine auxotrophy is fatal to engineered microbes. Namely, given the nature of our project and safeguards we have implemented, none of our BioBricks confer unsafe risks to individuals nor the environment.</span> | <li class="c0"><span class="c1">The engineered organism could potentially disrupt existing ecologies and be difficult to contain; however, as mentioned previously, integrated BBI would not make a difference in the bacteria’s survivability outside the lab, and we have taken care to ensure that the Threonine auxotrophy is fatal to engineered microbes. Namely, given the nature of our project and safeguards we have implemented, none of our BioBricks confer unsafe risks to individuals nor the environment.</span> | ||
</li> | </li> | ||
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<li class="c3">Our project utilized <span class="c1">Bacillus subtilis</span><span class="c1"> and a commonly used lab-strain of </span><span class="c8 c1">Escherichia coli</span><span class="c1"> TOP10 which are non-pathogenic and non-infectious as they are Biosafety Level 1 organisms (BSL-1). Thus the organisms were harmless to researchers. Additionally, the team worked with HCT116 cell lines, and 1BR3 cell lines which are Human colon carcinoma and Human skin fibroblast cell and are classified as Biosafety Level 2(BSL-2).The cell lines were received from completely anonymous donors. </span><span class="c4">We handled these cell lines at containment level 2 in accordance with the</span><span class="c1"> Bloodborne Pathogens Standard and Biosafety Committee</span><span class="c4"> guidelines.</span> | <li class="c3">Our project utilized <span class="c1">Bacillus subtilis</span><span class="c1"> and a commonly used lab-strain of </span><span class="c8 c1">Escherichia coli</span><span class="c1"> TOP10 which are non-pathogenic and non-infectious as they are Biosafety Level 1 organisms (BSL-1). Thus the organisms were harmless to researchers. Additionally, the team worked with HCT116 cell lines, and 1BR3 cell lines which are Human colon carcinoma and Human skin fibroblast cell and are classified as Biosafety Level 2(BSL-2).The cell lines were received from completely anonymous donors. </span><span class="c4">We handled these cell lines at containment level 2 in accordance with the</span><span class="c1"> Bloodborne Pathogens Standard and Biosafety Committee</span><span class="c4"> guidelines.</span> | ||
</li> | </li> |
Revision as of 02:17, 9 October 2016
Safety
Safety Considerations for Lab Work
All Principal Investigators, mentors, and undergraduate researchers were required to complete lab safety training and take safety courses developed by Environment Health and Safety (EHS) prior to working in the lab. The mandatory safety training included updated versions of the WHMIS course, the occupational health and safety course, the laboratory safety course, a hazard assessment course, an incident reporting and investigation course, a spill response course, a biosafety program course, a biosafety laboratory course, and a biosafety bloodborne pathogens course. The courses cover biological containment protocols, handling of hazardous materials such as liquid nitrogen, and disposal of waste as well as standard safety practices. All were required to take a test following each course, which certifies safe lab work under EHS Guidelines. All team members, advisors, and mentors received credit for each listed course and training program, and supervisors were present in the lab at all times to oversee undergraduate work.
Our project utilized BSL1 cells (E.coli and B.subtilis) that have GRAS labelling, thus our main project did not require ethics approval by the review boards. The University of Calgary has a university-wide Biosafety Committee, whose guidelines for safe biological laboratory practices were adhered to throughout the project.The team’s lab benches and experimental plans were assessed and deemed safe to proceed with. The Environment Health and Safety (EHS) provided training and information on the hazards from the types of sources and each form of radiation. All researchers underwent safety training courses. The individuals who worked with irradiated cells received radiation safety training from the EHS.
Our project utilized Bacillus subtilis and a commonly used lab-strain of Escherichia coli TOP10 which are non-pathogenic and non-infectious as they are Biosafety Level 1 organisms (BSL-1). Thus the organisms were harmless to researchers. Additionally, the team worked with HCT116 cell lines, and 1BR3 cell lines which are Human colon carcinoma and Human skin fibroblast cell and are classified as Biosafety Level 2(BSL-2).The cell lines were received from completely anonymous donors. We handled these cell lines at containment level 2 in accordance with the Bloodborne Pathogens Standard and Biosafety Committee guidelines.
Safety Considerations for Device
Patch Design
Choosing Patch Materials
Considering Human Use
Containment
The safety of the radioprotective patch was assessed by conducting in vivo mouse trails to test the patch, safety of the bacteria and the protein BBI. The mouse trial protocol was followed and approved by the Health Sciences Animal Care Committee (HSACC) who review animal care and testing.
- Link to certificate/Approval: .
- Documentation of ethics approval from the animal studies: available upon request
Future Considerations
A kill switch in Bacillus should be designed as engineering a kill switch into standardized plasmids could be useful for future iGEM competitions. Additionally, integrating BBI in multiple sites would give more auxotrophic sites, increasing the safety of using B.subtilis in the device.
Safe disposal:
- See pages 11 and 12 of the Device Manual for storage information.
Device Manual: https://drive.google.com/file/d/0B25REnfhyiNWM0hsNGtjRl9GWkk/view?usp=sharing
- Link to Safety forms:
- iGEM 2016 Basic Safety form
- Extended Safety Form Pt 1
- Extended Safety Form Pt 2