"
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 |
4. Do the biological materials used in your lab work pose any of the following risks? Please describe.
a. Risks to the safety and health of team members or others working in the lab?
b. Risks to the safety and health of the general public, if released by design or by accident?
c. Risks to the environment, if released by design or by accident?
d. Risks to security through malicious misuse by individuals, groups, or countries?
5. If your project moved from a small-scale lab study to become widely used as a commercial/industrial product, what new risks might arise? (Consider the different categories of risks that are listed in parts a-d of the previous question.) Also, what risks might arise if the knowledge you generate or the methods you develop became widely available? (Note: This is meant to be a somewhat open-ended discussion question.)
6. Does your project include any design features to address safety risks? (For example: kill switches, auxotrophic chassis, etc.) Note that including such features is not mandatory to participate in iGEM, but many groups choose to include them.
7. What safety training have you received (or plan to receive in the future)? Provide a brief description, and a link to your institution’s safety training requirements, if available.
8. Under what biosafety provisions will / do you work?
a. Please provide a link to your institution biosafety guidelines.
b. Does your institution have an Institutional Biosafety Committee, or an equivalent group? If yes, have you discussed your project with them? Describe any concerns they raised with your project, and any changes you made to your project plan based on their review.
c. Does your country have national biosafety regulations or guidelines? If so, please provide a link to these regulations or guidelines if possible.
d. According to the WHO Biosafety Manual, what is the BioSafety Level rating of your lab? (Check the summary table on page 3, and the fuller description that starts on page 9.) If your lab does not fit neatly into category 1, 2, 3, or 4, please describe its safety features [see 2013.igem.org/Safety for help].
e. What is the Risk Group of your chassis organism(s), as you stated in question 1? If it does not match the BSL rating of your laboratory, please explain what additional safety measures you are taking.
