Team:Kyoto/Safety

Safety

Q1. Would any of your project ideas raise safety issues in terms of: researcher safety, public safety, or environmental safety?

We use the following creatures and genes.

・Creatures

• Arabidopsis thaliana
• Escherichia coli

・Genes

• tatABCD operon
• TAMO reductase
• torA signal
• pspA
• Kil
• FLOWER LOCUS T
• others in iGEM Parts Kit

　FLOWER LOCUS T is derived from Arabidopsis thaliana and other all genes expect for iGEM Parts Kit are derived from E.coli. All experiments are conducted under the Biosafety level 1 control. All of rubbish containing recombinant E.coli is not carried out of our Lab before being sterilized. We always take care to lock the door of our Lab when there is no person so that the possibility that someone goes into our Lab and take away recombinant cells is very low.

　We are trained in applicable lab safety to ensure researcher safety and environmental safety. The researchers have also been trained in proper usage of chemicals and equipment. Some of the biological reagents and experiments necessary for the project needed safety measures. They were; Ethidium Bromide (EtBr), phenol, chloroform, 3,5-dinitrosalicylic acid, gas burner, ultra violet LED, polyacrylamide, 2-mercaptoethanol, PFA. Therefore, all researchers keep following rules when we use above chemicals or equipment.

• EtBr　(Ethidium bromide):　EtBr is regarded as a mutagen, carcinogen or teratogen. Lab members were briefed on the possible effects of EtBr. During the experiments, we wore gloves. After usage, the gloves and gel were separately disposed.
• Phenol and 3,5-Dinitrosalicylic Acid: Phenol and its vapors are corrosive to eyes, skin, and respiratory tract. 3,5-Dinitrosalicylic acid can cause serious irritation to eyes. These were handled under the draft chamber.
• Chloroform: Chloroform is a possible carcinogen. To avoid unnecessary exposure, this was also handled under the draft chamber.
• Gas Burner: We used gas burners and heat the air around the workspace to cause it to rise, thereby reducing contamination. Although the gas burner is a common combustion apparatus we are all familiar with, it can cause a massive disaster if used improperly. We made all-out efforts to keep all the flammable items away from the flame and not to pass behind a person who is using a gas burner.
• Ultra Violet LED: Ultra violet (UVB) is harmful to eyes. Hence, the LEDs were lit only inside the cardboard box and we only look at it to check if it is properly lighted to avoid long exposure.
• polyacrylamide,2-mercaptoethanol:We used these reagents for Western Blotting. They are toxic even though they are necessary to check the expression of target proteins so that we ask advice how we store them and obeyed the rules of Kyoto University.
• PFA: This reagents is used to microscope E.coli by Confocal laser scanning microscopy. This is ususally stored in Prof.Agata’s Laboratory

Q2. Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?

• NO. Currently we have no BioBrick which raises any safety issues. The BioBricks we made this year are derived only from lab-safe strains of Arabidopsis thaliana and E.coli.

Q3. Is there a local biosafety group, committee, or review board at your institution?

Yes. We submitted the experiment plan to the Environment, Safety, and Health Organization, Kyoto Univ. (http://www.esho.kyoto-u.ac.jp/index.php) and were allowed to operate genetic modification of the bacteria under the Biosafety level 1(. Experiments were planned following the safety guidelines of the university (http://www.esho.kyoto-u.ac.jp/wp-content /uploads/2008/05/25_01.pdf). These guidelines are based on national laws and this includes several regulations on GMOs (http://www.bch.biodic.go.jp/hourei1.html). We carried out our experiments in the students-laboratory of the Graduate School of Science / Faculty of Science, Kyoto Univ, which has its own department for safety and environment. All of the lab members received training for PCR, culture of cells, miniprep and etc., the minimal genetical operations and use of autoclave.

Q4. 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?

IGEMers should set up a common clear biosafety regulation in iGEM competition and establish a systematic guideline of education about biosafety. This education should include information about biohazard and biotechnology risk assessment and risk analysis.

　Backgrounds of iGEMers become wider and wider, from high school students to graduate student in biology and from major in physics to major in molecular biology. For some students, iGEM are the first contact to genetic engineering and have little knowledge of biohazard and importance of biosafety. Each iGEM teams, of course, teach them to their members. However, a systematic education way of biosafety will help students to understand biosafety and to conduct their experience in adequate ways.

From these reason, we recommend that iGEM establish a systematic guideline of education about biosafety.