Team:Freiburg/Safety/safety forms

From 2013.igem.org

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Revision as of 15:00, 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

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