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Team:Freiburg/Safety/safety forms
From 2013.igem.org
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.
