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From 2012.igem.org

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Safety Questions:

1. Would any of your project ideas raise safety issues in terms of: researcher safety, public safety, or environmental safety?

a) First of all, we have ensured the safety of the researchers. The safety regulations in the laboratory and be strictly obeyed. There is an isolated area for us to experiment when we have to use toxic chemicals as EB or other material. And the laboratory where we have been doing our work has staff to reduce the potential safety hazard.

b) Second, we have ensured the public safety by doing our experiment within the area of our laboratory strictly. And of course, the laboratory building where we have been working has a specific safety system for garbage disposal or recycle the garbage.

c) The third, we have also considered the environmental safety. Besides the effort we have done above, the gene part we chose has little potential to raise safety issues to environment. And the materials we have used are not allowed to be taken out of the laboratory.

2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?

a) No. On one hand, almost all the Biobrick parts we used for iGEM are from the DNA kit, and the plasmids in the kit are safe due to the well-developed technique of transfection and screening though they are carrying the antibiotic resistance genes. On the other hand, our own Biobrick and strain we used has little potential to risk the environment.

3. Is there a local biosafety group, committee, or review board at your institution?

a) Yes. The State Key Laboratory of Bio-control is located in our school, and most of our experiments are done in a subsidiary lab of it. Although our project will not course any serious biosafety problem, we check each part and material used for the project carefully and all are approved safe by that state key lab.

4. 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?

a) We are trying to use Plasmid Addition genes to avoid or cut down the possibilities about three biosafety core problems: Horizontal Gene Transfer (HGT), foreign genes’ unstability(or the parts, devices, and systems) and antibiotic using.

b) Plasmid Addiction

i. They are come from natural plasmid systems (e.g. Plasmid F from E.coli), which contains a pair of stable toxin and an unstable antidote. The genes can both express when the plasmid is in the cell. If the plasmid is missing by accident, the unstable antidote is decomposed by some enzymes, and the toxin causes the plasmid-missing bacteria to die.

c) Device Construction

i. There two proper pairs of toxin and antidote, named the Pair 1 (ccdB and ccdA from E.coli`s F plasmid )and Pair 2（ParE and ParD from E.coli`s plasmid）

ii. Plasmid Vector: Insert Toxin 1 gene and Antidote 2 gene in the Plasmid vector.

iii. Bacteria Genome: Insert Toxin 2 gene and Antidote 1 gene in the bacteria genome through transposon.

iv. When HGT happens, due to the missing of the Antidote 1 gene and its production in the new host, the Toxin 1 will cause the new host to die.

v. When the plasmid vector is missing, the missing of the Antidote 2 will cause the cell to die. And statistically, the intact host cells will be the overwhelming majority. And the Plasmid Addition is established.

vi. The toxin`s function can replace the antibiotic selection.

vii. We will choose as many pairs as we could and use mathematical modeling to model the system and optimize the efficiency of the different combinations.