Safety Proposal

Recently, the outbreak of the new and fatal form of E.coli has awakened the public awareness on the safety of handling bacteria. This makes us design our safety strategy more seriously and thoroughly. When it comes to laboratory work, safety should always be put in the first priority. Otherwise, the laboratory practice could never be sustainable. Furthermore, not only the individual safety of the researchers but also the public, the environmental safety, and even the safety issues for the future iGEM teams, are concerned.

A. Safety issues of our Projects

Our project needs to insert a light-driven ion pump, halorhodopsin, into the magnetotactic bacteria. Thus, the bacteria can uptake chloride ions into the cells and decrease the salinity of the environment. By applying magnetic field, we can control the movement of bacteria and gives out a salinity gradient. We will then manufacture a pair of electrodes in order to generate electricity by combining sunlight, salinity difference together with the genetically engineered bacteria. Regarding to our project, we considered several safety issues that possibly be raised.

1. General biological safety

With regard to our researchers' safety, we have several precautions to ensure our team members can work in a safe condition. First and foremost, researchers have to attend laboratory safety workshops hold by University Safety and Environment Office before they could join any laboratory work. The workshops cover mainly 4 safety aspects, including general, chemical, radiation and biological safety. Also, the researchers have to wear gloves and laboratory coats throughout the laboratory work to avoid contacting any harmful, irritant, toxic or even carcinogenic reagents. Moreover, our project as well as the laboratory work is supervised by three professors together with three instructors who got Biological safety level Ⅱ certificate. Our researchers have also received biological safety level I certificate. Furthermore, our laboratory is a registered student laboratory with level Ⅱ safety which is suitable for bacterial and cell culture use. Inside the laboratory, students operate with autoclaved materials and follow cleaning procedures every time. We clean benches with 70% ethanol before and after we perform experiments. Last but not least, all the students maintain a good biological safety hood throughout the project in order to reduce any risk factor to the minimum.

2. Public and Environmental Safety

Firstly, we followed the regulation of the Hong Kong Government to purchased bacteria. Regarding to the bacteria we used, we chose 2 non-virulent strains of E.coli: DH5a for amplifying BioBrick and BL21 for expression of protein. To add on, Magnetospirillum gryphiwaldense MSR-1, Halobacterium salinarum, Haloterrigena turkmenica and Natronomonas pharaonis that we manipulated are all non-virulent. These are to avoid putting public and environmental safety at risk. We follow safety rules so as to prevent no harmful gene from leakage. Genomic DNA has also been tested and verified before use. And we disinfect all the cultures and wastes by autoclaving and bleaching before disposal to avoid leakage of any other potential risky substances e.g. genetically engineered bacteria. Finally, we promote synthetic biology and BioBrick to the public so more people can get familiar to this research area, and understand the importance of research safety. Through combining the above methods, therefore, we believe our project would neither cause any harm to the public nor the environment.

3. BioBricks

To answer the question "Do any of the new BioBrick parts (or devices) that you made this year raise any safety issues?" We believe our answer is No, the BioBrick parts we made this year are not going to raise any safety issues. The halorhodopsin ion pump and magnetosome genes that we focused on exist in the nature. Both of them are observed not to be infectious and pathogenic. So, the BioBricks we are going to make this year are not risky.

4. Antibiotics resistant plasmid

There is a concern that using antibiotic containing plasmid for transformation selection may possibly lead to the production of drug-resistance bacteria. The way of avoiding leakage of genetically engineered bacteria should also be considered. We make sure that every time we dispose bacterial culture, we process them with 30% bleach together with autoclaving procedures to try to reduce the probability of leakage to the minimum. Moreover, we transform bacteria with one single type of antibiotic every time, so as to ensure bacteria can only achieve resistance to one kind of . Therefore, using other types of antibiotic can still kill the bacteria to stop the spreading.

5. Generating Eletrodes

Firstly, all our team members involved in handling of fine powder, such as graphite and sodium manganese oxide, are required to wear surgical or N95 mask in order to avoid the risk of irritation. Also, the team member responsible for the key steps in making the electrode has received formal training for working in the Nano Fabrication Laboratory and has a good understanding of the potential risks involved. Furthermore, we noticed that there is a small risk of cut involved in fabricating the electrodes into desired shape, where sharp tools like jigsaw may be used. In response to this, we decided at least one other person must be present for surveillance and reminding the working person. And a tidy working environment with good lighting and first-aid kit readily available will be provided for mechanical work.

6. Electric Circuit

There are common potential risks for short-circuiting, overheating and electric shock when it comes to using electric circuits. With regard to this, the circuit that we chose is equipped with modern safety mechanisms such as overload, overhear as well as short-circuit protection. To add on, the production of electric circuits involves the use of harmful chemicals, risk of burns in soldering, and cuts due to sharp points at the bottom of the circuit board. However, the team members responsible for designing and fabricating the electric circuits have received professional training in this aspect, and have prior experience in making electric circuit.

B. Local Biosafety Group at the Institution

The Chinese University of Hong Kong has a specific group, named University Safety & Environment Office. The office works for guidelines to all the faculties for safety issues, for example, laboratory safety, public safety, etc. Special guidelines which we strictly follow are given to us for handling microorganisms in the laboratory.

C. Suggestions to future iGEM teams

We suggest future iGEM teams not to manipulate any infectious or virulent bacteria in the project, so as to avoid any chance of causing harm because of executing the project idea. Also, we advice we should not use any harmful gene in the project. As chances of gene leakage are not possible to reduce to zero, harmful genes should be avoided.