Difference between revisions of "Team:UC Davis/Safety"

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Click <a href="https://2018.igem.org/File:T--UC_Davis--SDS.xlsx">here</a> to view our submitted safety spreadsheet.
 
Click <a href="https://2018.igem.org/File:T--UC_Davis--SDS.xlsx">here</a> to view our submitted safety spreadsheet.
  
<p>We are dedicated to following the safety and security rules of our university and the iGEM competition, as well as applicable state and national standards. Our efforts include designing an experiment which minimizes risk and exposure to hazardous substances and cells, working in an appropriate setting, and being properly trained by safety officers at our university.  
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<p>We are dedicated to following the safety and security rules of our university and the iGEM competition, as well as applicable state and national standards. As part of our project, we also investigated how our project might fit into local regulations which have more strict restrictions on biotechnology than the prevailing national standards. For more information regarding this, please see our Guide to Integrated Human Practices.  
 
</p>
 
</p>
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<p> We have taken steps to design our experiment in such a way which minimizes risk and exposure to hazardous substances and cells by working in an appropriate setting, and being properly trained by safety officers at our university.
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Project Overview
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  Our project aims to develop mammalian-based biosensors for use in the context of environmental toxicology. Specifically, out biosensors have been designed to co-opt the mammalian cell's intrinsic stress response pathways and use these to trigger the production of a fluorescent reporter. We hypothesize that a device reporting on the activation of cell stress pathways will provide more physiologically and health relevant information about the potential toxins present in an environment than bioassays which seek to simply measure compound presence and/or abundance. That is, our bioassay asks not whether a compound is there, but rather whether compounds exists that may pose a health hazard. We leave the identification of the specific hazards to more sensitive and quantitative techniques.
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<p>Our biosensors use mammalian-derived promoters of genes known to be activated in response to stress-inducing environmental pollutants. These promoters are coupled to a reporter gene (eGFP) and used in in vitro assays to report on the presence of compounds that elicit cell stress.
 
 
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<p>The specific promoters we have identified for our project control the expression of the following genes: Metallothionein 1, Metallothionein 2, Cytochrome P450(CYP1A1), Fibroblast growth factor(FGF21), and the Growth arrest and DNA Damage(GADD45a and GADD153). The representative environmental toxins that we plan to stimulate our engineered cells with include: 2,4-Dichlorophenoxyacetic acid, Warfarin, Metam Sodium, and Copper Sulfate. We are also using Hydrogen Peroxide as a positive control.
 
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   Lab Safety Training
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   Lab Safety Training and Institutional Requirements
  
 
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   All Team members have taken in-person Laboratory Safety training for Biosafety, Chemical Safety, Radiation Compliance, Bloodborne Pathogens, Medical Waste, and Laboratory Ergonomics. All team members are also certified to use the Biological Safety Cabinet to work with BSL1 and BSL2 mammalian cell lines (Only using BSL1 for this project).  
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   Before work began, all team members were given site specific safety training and a general safety training course for work in a molecular biology lab. Additionally, all Team members have taken in-person Laboratory Safety training for Biosafety, Chemical Safety, Radiation Compliance, Bloodborne Pathogens, Medical Waste, and Laboratory Ergonomics. All team members are certified to use the Biological Safety Cabinet and to work in BSL1 and BSL2 laboratories (although we only required BSL1 to complete this project). Our campus Biological Use Authorization (BUA) committee has reviewed and approved our project involving work with the mammalian cell lines, CHO DG-44 and AML12. We were granted UC Davis BUA #R2565.
<p>Our campus Biological Use Authorization committee has reviewed and approved our project and work with CHO DG-44 and AML12 cells in the lab. Our UC Davis BUA# is: R2565
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   We are working laboratory grade Warfarin, Hydrogen Peroxide, 2,4-D, Metam Sodium, and Copper Sulfate.  
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   The representative environmental toxins that we exposed our bioassay to were 2,4-dichlorophenoxyacetic acid <a href="https://www.sigmaaldrich.com/MSDS/MSDS/PleaseWaitMSDSPage.do?language=&country=US&brand=ALDRICH&productNumber=CDS005520&PageToGoToURL=https%3A%2F%2Fwww.sigmaaldrich.com%2Fcatalog%2Fsearch%3Fterm%3D2%252C4-D%26interface%3DAll%26N%3D0%26mode%3Dpartialmax%26lang%3Den%26region%3DUS%26focus%3Dproduct">(SDS)</a>, warfarin <a href="https://www.sigmaaldrich.com/MSDS/MSDS/DisplayMSDSPage.do?country=US&language=en&productNumber=258016&brand=ALDRICH&PageToGoToURL=https%3A%2F%2Fwww.sigmaaldrich.com%2Fcatalog%2Fproduct%2Faldrich%2F258016%3Flang%3Den">(SDS)</a>, and copper sulfate <a href="https://www.sigmaaldrich.com/MSDS/MSDS/DisplayMSDSPage.do?country=US&language=en&productNumber=H1009&brand=SIGMA&PageToGoToURL=https%3A%2F%2Fwww.sigmaaldrich.com%2Fcatalog%2Fproduct%2Fsigma%2Fh1009%3Flang%3Den">(SDS)</a>. Additionally, we used hydrogen peroxide <a href="https://www.sigmaaldrich.com/MSDS/MSDS/DisplayMSDSPage.do?country=US&language=en&productNumber=H1009&brand=SIGMA&PageToGoToURL=https%3A%2F%2Fwww.sigmaaldrich.com%2Fcatalog%2Fproduct%2Fsigma%2Fh1009%3Flang%3Den">(SDS)</a> as a positive control. Click <a href="https://2018.igem.org/File:T--UC_Davis--spillresponse.pdf"> here</a> for safety measures regarding the spill of hazardous chemicals. Warfarin (GHS code H360) is a reproductive toxin. Click <a href="https://2018.igem.org/File:T--UC_Davis--reproductivetoxin.pdf">here</a> for the SOP used when handling this chemical. Chemicals were appropriately disposed through the guidelines set by the UC Davis WASTe system. We also collected an environmental sample of water from a tributary of Putah Creek, from Davis, California. This environmental sample was autoclaved and UV sterilized to kill any pathogens present, before working with the sample.
<p> Click <a href="https://2018.igem.org/File:T--UC_Davis--spillresponse.pdf">here</a> for safety measures regarding the spill of hazardous chemicals.
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</p>
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<p>Additionally, Warfarin (GHS code H360) is a reproductive toxin. Click <a href="https://2018.igem.org/File:T--UC_Davis--reproductivetoxin.pdf">here</a> for the SOP used when handling this chemical.
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</p>
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<p>Chemicals are appropriately disposed of through guidelines set by the UC Davis WASTe system.
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   Our project involves Biosafety level one work. The organism and cell lines we are using pose little risk to us, our colleagues, the community or the environment. These risks are further minimized by adhering to the administrative and engineered safety controls in the lab. Administratively, we were all required to take the standard our campus wide safety training, in person site specific training, and additional training pertaining to handling and safe practices with working with BSL-1 organisms and cell lines (see the list of training courses below). Personal protective equipment is used whenever working with these organisms and strains in the lab. All items in contact with E. coli and the cell lines are sterilized by autoclave and/or bleach (final concentration: 20%, 20 minute contact time) prior to disposal. Work with BSL-1 cell lines are restricted to the biosafety cabinet in the lab.  
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   Our project involves Biosafety Level One work. The organism and cell lines we are using pose little risk to us, our colleagues, the community or the environment. These risks are further minimized by adhering to the administrative and engineered safety controls in the lab. Administratively, we were all required to take the standard our campus wide safety training, in person site specific training, and additional training pertaining to handling and safe practices with working with BSL-1 organisms and cell lines (see the list of training courses below). Personal protective equipment is used whenever working with these organisms and strains in the lab. All items in contact with E. coli and the cell lines are sterilized by autoclave and/or bleach (final concentration: 20%, 20 minute contact time) prior to disposal. Work with BSL-1 cell lines are restricted to the biosafety cabinet in the lab.
  
<p>The threats posed by the CHO and AML cells are minimal. Both our cell lines have been categorized as non-infectious and non-toxic. Mammalian cells are optimized to live inside of mammals, and are not likely to survive in the natural environment or infect other mammals.
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<p> The threats posed by the CHO and AML cells are minimal. Both our cell lines have been categorized as non-infectious and non-toxic. Mammalian cells are optimized to live inside of mammals, and are not likely to survive in the natural environment or infect other mammals.
 
</p>
 
</p>
 
<p>In order to prevent situations where the chassis organisms escape the lab, all members present in the lab space follow the safety regulations detailed by UC Davis. As an additional precaution all strains are sterilized via bleach (final concentration: 20%, 20 minute contact time) and/or autoclaving before disposal into the general garbage stream.
 
<p>In order to prevent situations where the chassis organisms escape the lab, all members present in the lab space follow the safety regulations detailed by UC Davis. As an additional precaution all strains are sterilized via bleach (final concentration: 20%, 20 minute contact time) and/or autoclaving before disposal into the general garbage stream.
 
</p>
 
</p>
<p>We will also be using E. coli in our experiments to replicate DNA. We are only going to be using non-pathogenic strains of E. coli. E. coli is not permitted to leave the lab and is sterilized via bleach (final concentration: 20%, 20 minute contact time) and/or autoclave prior to disposal.
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<p>We used E. coli (non-pathogenic strain DH5α) to clone DNA. E. coli was not permitted to leave the lab and was sterilized via bleach (final concentration: 20%, 20 minute contact time) and/or autoclave prior to disposal.
 
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   Our project was intended only to be used in a laboratory setting. This use involves minimal risks to safety, security, or ethics. Our anticipated real world application is carried out in a laboratory setting, and the safety protocols that will be followed are the same as the ones we are currently following.  
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   Our project was intended only to be used in a laboratory setting. We never plan on releasing our transgenic cell lines into the environment. Our anticipated real world application is carried out in a laboratory setting, and the safety protocols that will be followed are the same as the ones we are currently following. We investigated a variety of ethical considerations and how our project might fit into different cultural contexts, and documented these investigations in our Guide to Integrated Human Practices document.  
  
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  Security and Dual Use Risks
  
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  We evaluated our project to have minimal security risks. We did not work with any pathogens or parts originating in pathogens, nor did we produce any biological products which may have security risks associated with them.
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Revision as of 20:27, 2 October 2018

iGEM

Safety
Safety
We submitted our final iGEM safety form by the deadline on September 7th and it has been approved. Click here to view our submitted safety spreadsheet.

We are dedicated to following the safety and security rules of our university and the iGEM competition, as well as applicable state and national standards. As part of our project, we also investigated how our project might fit into local regulations which have more strict restrictions on biotechnology than the prevailing national standards. For more information regarding this, please see our Guide to Integrated Human Practices.

We have taken steps to design our experiment in such a way which minimizes risk and exposure to hazardous substances and cells by working in an appropriate setting, and being properly trained by safety officers at our university.

Lab Safety Training and Institutional Requirements
Before work began, all team members were given site specific safety training and a general safety training course for work in a molecular biology lab. Additionally, all Team members have taken in-person Laboratory Safety training for Biosafety, Chemical Safety, Radiation Compliance, Bloodborne Pathogens, Medical Waste, and Laboratory Ergonomics. All team members are certified to use the Biological Safety Cabinet and to work in BSL1 and BSL2 laboratories (although we only required BSL1 to complete this project). Our campus Biological Use Authorization (BUA) committee has reviewed and approved our project involving work with the mammalian cell lines, CHO DG-44 and AML12. We were granted UC Davis BUA #R2565.
Chemical Safety
The representative environmental toxins that we exposed our bioassay to were 2,4-dichlorophenoxyacetic acid (SDS), warfarin (SDS), and copper sulfate (SDS). Additionally, we used hydrogen peroxide (SDS) as a positive control. Click here for safety measures regarding the spill of hazardous chemicals. Warfarin (GHS code H360) is a reproductive toxin. Click here for the SOP used when handling this chemical. Chemicals were appropriately disposed through the guidelines set by the UC Davis WASTe system. We also collected an environmental sample of water from a tributary of Putah Creek, from Davis, California. This environmental sample was autoclaved and UV sterilized to kill any pathogens present, before working with the sample.
Environmental Safety
Our project involves Biosafety Level One work. The organism and cell lines we are using pose little risk to us, our colleagues, the community or the environment. These risks are further minimized by adhering to the administrative and engineered safety controls in the lab. Administratively, we were all required to take the standard our campus wide safety training, in person site specific training, and additional training pertaining to handling and safe practices with working with BSL-1 organisms and cell lines (see the list of training courses below). Personal protective equipment is used whenever working with these organisms and strains in the lab. All items in contact with E. coli and the cell lines are sterilized by autoclave and/or bleach (final concentration: 20%, 20 minute contact time) prior to disposal. Work with BSL-1 cell lines are restricted to the biosafety cabinet in the lab.

The threats posed by the CHO and AML cells are minimal. Both our cell lines have been categorized as non-infectious and non-toxic. Mammalian cells are optimized to live inside of mammals, and are not likely to survive in the natural environment or infect other mammals.

In order to prevent situations where the chassis organisms escape the lab, all members present in the lab space follow the safety regulations detailed by UC Davis. As an additional precaution all strains are sterilized via bleach (final concentration: 20%, 20 minute contact time) and/or autoclaving before disposal into the general garbage stream.

We used E. coli (non-pathogenic strain DH5α) to clone DNA. E. coli was not permitted to leave the lab and was sterilized via bleach (final concentration: 20%, 20 minute contact time) and/or autoclave prior to disposal.

Ethical Risks
Our project was intended only to be used in a laboratory setting. We never plan on releasing our transgenic cell lines into the environment. Our anticipated real world application is carried out in a laboratory setting, and the safety protocols that will be followed are the same as the ones we are currently following. We investigated a variety of ethical considerations and how our project might fit into different cultural contexts, and documented these investigations in our Guide to Integrated Human Practices document.
Security and Dual Use Risks
We evaluated our project to have minimal security risks. We did not work with any pathogens or parts originating in pathogens, nor did we produce any biological products which may have security risks associated with them.
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UC Davis iGEM 2018