Team:Freiburg/Safety/safety forms

From 2013.igem.org

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<p id="h3"> 2. Highest Risk Group Listed </p>
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<p id="h3">Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues? If yes, </p>
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<p>[ ]1 <br>
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[2] Greater than 1 </p>
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<ul>
 
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<li><b>did you document these issues in the Registry? </b></li>
 
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<li><b>how did you manage to handle the safety issue?</b> </li>
 
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<li><b>how could other teams learn from your experience? </b></li>
 
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</ul>
 
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<p>
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<p id="h3"> 3. List and describe all new or modified coding regions you will be using in your project. (If you use parts from the 2013 iGEM Distribution without modifying them, you do not need to list those parts.) </p>
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Our constructs do not raise any safety issues. They should not be able to increase or give pathogenicity to the applied microorganisms or cell lines. The single parts do not encode for any toxins. The devices as they are used in our project should also represent no hazard for public health or environmental safety. In their current state misuse according to bioterrorism should also be unlikely.
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</p>
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<p id="h3">Is there a local biosafety group, committee, or review board at your institution? </p>
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<table id="tabelle">
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<table border="3" frame="box">
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<ul>
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<tr> <th> Part number. </th> <th> name </th> <th> "Where did you get the
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<li><b>If yes, what does your local biosafety group think about your project? </b></li>
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physical DNA for this
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</ul>
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part (which lab,
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synthesis company, etc)
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" </th> <th> "What species does
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this part originally
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come from?
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" </th> <th> "What is the
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Risk Group of
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the species?
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" </th> <th> "What is the function of
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this part, in its parent
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species?
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" </th> </tr>
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<tr> <td> BBa_K1150000 </td> <td> cas9 </td> <td> AddGene </td> <td> Streptococcus pyogenes </td> <td> 2 </td> <td> specific immune response </td> </tr>
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<tr> <td> BBa_K1150001 </td> <td> vp16 </td> <td> AG Wilfried Weber, University of Freiburg </td> <td> Herpes simplex virus </td> <td> 2 </td> <td> gene transcription stimulator </td> </tr>
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<tr> <td> BBa_K1150002 </td> <td> krab </td> <td> AG Wilfried Weber, University of Freiburg </td> <td> Homo sapiens </td> <td> 1 </td> <td> repressor of transcriptional activity </td> </tr>
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<tr> <td> BBa_K1150003 </td> <td> g9a-sd </td> <td> AG Jeltsch, University of Stuttgart </td> <td> Mus musculus </td> <td> 1 </td> <td> histone methyl transferase </td> </tr>
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<tr> <td> BBa_K1150004 </td> <td> phyb </td> <td> AG Wilfried Weber, University of Freiburg </td> <td> Arabidopsis thaliana </td> <td> 1 </td> <td> light signaling transducer </td> </tr>
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<td> BBa_K1150005 </td> <td> pif6 </td> <td> AG Wilfried Weber, University of Freiburg </td> <td> Arabidopsis thaliana </td> <td> 1 </td> <td> interacting factor of phyB </td> </tr>
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<td> BBa_K1150006 </td> <td> uvr8 </td> <td> AG Wilfried Weber, University of Freiburg </td> <td> Arabidopsis thaliana </td> <td> 1 </td> <td> light signaling transducer </td> </tr>
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<td> BBa_K1150007 </td> <td> cop1 </td> <td> AG Wilfried Weber, University of Freiburg </td> <td> Arabidopsis thaliana </td> <td> 1 </td> <td> light signaling transducer </td> </tr>
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<td> BBa_K1150008 </td> <td> cip </td> <td> AG Wilfried Weber, University of Freiburg </td> <td> Arabidopsis thaliana </td> <td> 1 </td> <td> light signaling transducer </td> </tr>
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<td> BBa_K1150009 </td> <td> cry2 </td> <td> AG Wilfried Weber, University of Freiburg </td> <td> Arabidopsis thaliana </td> <td> 1 </td> <td> light signaling transducer </td> </tr>
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<td> BBa_K1150010 </td> <td> NLS </td> <td> AG Wilfried Weber, University of Freiburg </td> <td> AAV2 </td> <td> 2 </td> <td> nuclear localization sequence </td> </tr>
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<td> BBa_K1150011 </td> <td> SV40 </td> <td> AG Wilfried Weber, University of Freiburg </td> <td> Simian-Virus 40 </td> <td> 2 </td> <td> viral promoter </td> </tr>
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<td> BBa_K1150012 </td> <td> bGH-terminator </td> <td> AG Wilfried Weber, University of Freiburg </td> <td> Bos taurus </td> <td> 1 </td> <td> terminator of the bovine growth hormone gene </td> </tr>
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<td> BBa_K1150013 </td> <td> short linker </td> <td> AG Wilfried Weber, University of Freiburg </td> <td> synthesized by Sigma-Aldrich as Oligos </td> <td> </td> <td> short, flexible linker </td> </tr>
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<td> BBa_K1150015 </td> <td> CMV </td> <td> AG Wilfried Weber, University of Freiburg </td> <td> Cytomegalievirus </td> <td> 2 </td> <td> viralpromoter </td> </tr>
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<td> BBa_K1150016 </td> <td> HA-Tag </td> <td> AG Wilfried Weber, University of Freiburg </td> <td> synthesized by Sigma-Aldrich as Oligos </td> <td> </td> <td> Protein sequence tag </td> </tr>
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<td> BBa_K1150034 </td> <td> RNA-plasmid </td> <td> AddGene </td> <td> Streptococcus pyogenes, Homo sapiens </td> <td> 2 </td> <td> specific immune response </td> </tr>
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</table>

Revision as of 14:57, 1 October 2013

Safety questions

At the beginning of our research we wanted to be aware of all the probable hazards concerning our project. This included that we tried to identify safety issues. Therefore we concentrated on pathogenicity of the microorganisms and cell lines of interest, the datasheets of chemicals probably used during the project (e.g. DNA stains) and the engineered devices and systems. Thus, we orientated on the hints of the iGEM 2013 safety page as well as the safety constraints for genetic engineering given by the “Stabsstelle Sicherheit” of the University of Freiburg. At this point we are obliged to Dr. M. Zurbriggen, who gave us safety instructions before starting our investigations.

1.Please describe the chassis organism(s) you will be using for this project. If you will be using more than one chassis organism, provide information on each of them:

# Species Strain no/name Risk Group Risk group source link Disease risk to humans? If so, which disease?
1 E.coli (K12) TOP10 1 http://apps2.bvl.bund.de/strainwww/protected/main/strain.do?method=detail&theId=49&d-49653-p=null "Yes. May cause irritation to skin, eyes, and respiratory tract, may affect kidneys. "
2 human HEK293T 2 (1, according to german guidelines) http://apps2.bvl.bund.de/cellswww/protected/main/cell.do?method=detail&theId=73&d-49653-p=22
3 human HeLa 2 (1, according to german guidelines) http://apps2.bvl.bund.de/cellswww/protected/main/cell.do?method=detail&theId=22&d-49653-p=null
4 hamster CHO-K1 1 http://apps2.bvl.bund.de/cellswww/protected/main/cell.do?method=detail&theId=13&d-49653-p=12
5 mouse NIH/3T3 1 http://apps2.bvl.bund.de/cellswww/protected/main/cell.do?method=detail&theId=33&d-49653-p=null

2. Highest Risk Group Listed

[ ]1
[2] Greater than 1

3. List and describe all new or modified coding regions you will be using in your project. (If you use parts from the 2013 iGEM Distribution without modifying them, you do not need to list those parts.)

Part number. name "Where did you get the physical DNA for this part (which lab, synthesis company, etc) " "What species does this part originally come from? " "What is the Risk Group of the species? " "What is the function of this part, in its parent species? "
BBa_K1150000 cas9 AddGene Streptococcus pyogenes 2 specific immune response
BBa_K1150001 vp16 AG Wilfried Weber, University of Freiburg Herpes simplex virus 2 gene transcription stimulator
BBa_K1150002 krab AG Wilfried Weber, University of Freiburg Homo sapiens 1 repressor of transcriptional activity
BBa_K1150003 g9a-sd AG Jeltsch, University of Stuttgart Mus musculus 1 histone methyl transferase
BBa_K1150004 phyb AG Wilfried Weber, University of Freiburg Arabidopsis thaliana 1 light signaling transducer
BBa_K1150005 pif6 AG Wilfried Weber, University of Freiburg Arabidopsis thaliana 1 interacting factor of phyB
BBa_K1150006 uvr8 AG Wilfried Weber, University of Freiburg Arabidopsis thaliana 1 light signaling transducer
BBa_K1150007 cop1 AG Wilfried Weber, University of Freiburg Arabidopsis thaliana 1 light signaling transducer
BBa_K1150008 cip AG Wilfried Weber, University of Freiburg Arabidopsis thaliana 1 light signaling transducer
BBa_K1150009 cry2 AG Wilfried Weber, University of Freiburg Arabidopsis thaliana 1 light signaling transducer
BBa_K1150010 NLS AG Wilfried Weber, University of Freiburg AAV2 2 nuclear localization sequence
BBa_K1150011 SV40 AG Wilfried Weber, University of Freiburg Simian-Virus 40 2 viral promoter
BBa_K1150012 bGH-terminator AG Wilfried Weber, University of Freiburg Bos taurus 1 terminator of the bovine growth hormone gene
BBa_K1150013 short linker AG Wilfried Weber, University of Freiburg synthesized by Sigma-Aldrich as Oligos short, flexible linker
BBa_K1150015 CMV AG Wilfried Weber, University of Freiburg Cytomegalievirus 2 viralpromoter
BBa_K1150016 HA-Tag AG Wilfried Weber, University of Freiburg synthesized by Sigma-Aldrich as Oligos Protein sequence tag
BBa_K1150034 RNA-plasmid AddGene Streptococcus pyogenes, Homo sapiens 2 specific immune response

Yes, among other things, the department “Stabsstelle Sicherheit” is responsible for questions and issues concerning biosafety at the University of Freiburg.

Do you have any other ideas how to deal with safety issues that could be useful for future iGEM competitions? How could parts, devices and systems be made even safer through biosafety engineering?

In our lab, we were especially concerned with carcinogenity. To protect our team we banned ethidium bromide completely and used next generation DNA stains concerned to be less carcinogenic. Additionally, nitrile gloves were used while cutting agarose gels containing DNA intercalating substances. Another hazard is the UV light, which can lead to mutations in DNA. Eyes and skin were protected due to lab coats and UV shield helmets. Theses safety precautions proved themselves in practice and are recommended to all other iGEM-teams.

Systems can be made safer through genetically switches. Our constructs are induced or silenced by stimuli, e.g. light of special wavelengths or hormones. Without the right stimulus the system is not running. Other possibilities are constructs that regulate themselves or self destruction mechanisms in case of dysfunctions.