Any scientific research project involves in the field of life science needs to be carried out healthily under the guidelines of biosafety, biosafety and bioethics. Especially under the rapid development of the emerging discipline of synthetic biology, any behavior beyond the minefield will cause a strong social shock. At this time, young iGEMers should have the courage to assume the responsibility of maintaining biosafety, and always pay attention to safety issues in the process of doing experiments. Therefore, this year, our team conducted an analysis of possible biosafety risks and laboratory safety risks, and lists the following two aspects.
Microorganisms are classified into four Risk Groups：
Risk Group 1 : do not cause disease in healthy adult humans.
Risk Group 2 : can cause disease in humans, but the disease is treatable or preventable.
Risk Group 3 ：cause serious disease in humans. Treatments and vaccines for these diseases may exist.
Risk Group 4 ：cause deadly disease in humans, and they can easily travel from one person to another. No treatments or vaccines exist for these diseases.
Our project uses the CRISPR-Cas12a gene editing system to genetically modify Corynebacterium glutamicumATCC13032 to achieve high levels of lycopene production.The microbe used in the project: Corynebacterium glutamicum ATCC13032 and Escherichia coli Top10. These two microorganisms have a safety rating of level 1 and belong to the iGEM white list for safety.
Our project is based on the metabolic engineering of Corynebacterium glutamicum to express lycopene. To increase lycopene production, some genes need to be inserted into the microbe. All of these genes are derived from the genome of Corynebacterium glutamicumitself, so these genes have a high safety profile.
Before the experiment: Before the experimenter, the experimenter should wear experimental clothes, tie up their long hair, and wear trousers and non-toe shoes.
Experimental: Gloves should be worn when contact with toxic reagents and organic reagents. And masks should be worn when contact with pungent reagents.
End of the experiment: Arrange everything before leaving the lab and wash your hands before leaving. The last person to leave the laboratory should check whether the taps are turned off.
The opening date for all enzymes, antibodies and chemical reagents must be recorded.
The liquid shall not exceed 2 / 3 of the volume; otherwise, it may cause gushing or blowout, thus contaminate surrounding samples or injure personnel.
Place the reagent in the right place, label it properly and mark it properly.
Do not bring contact with molecular dyes (Gelview) to the usual place for experiments.
Regularly check whether the instrument is available, if there is a problem, the professional needs to come to repair.
The water bath should be closed in time after use.
When using the gel imaging system, it should be careful not to open the front panel of the system during the ultraviolet light source photography. If the glue cutting recovery operation is required, it should wear latex gloves and PE gloves to minimize the impact. Remove the gel and turn off the instrument after use.
Use the centrifuge to balance. After the centrifuge, wait until the speed is reduced to 0 r/min before opening the centrifuge.
Before using the sterilization pot, add an appropriate amount of DI water to the pot, and put the sterilized items into the basket and then put them into the sterilization pot smoothly. After sterilization, open the lid until the temperature drops below 90°C and the pressure is 0 MPa.
Before operating on the ultra-clean workbench, you should open the ultraviolet sterilization in advance. During the sterilization period, people can not directly contact ultraviolet light. After use, the clean table should turn off the alcohol lamp and turn off the lighting and ventilation.
The location of the same class of chemicals and appropriate storage methods, such as primers, enzymes, plasmids, strains, antibodies and marker were stored in -20°C refrigerators, prepared competent cells in -80°C refrigerators. The molecular dye (Gelview) shall be placed alone on the console dedicated for gel electrophoresis.
The iGEM HBUT-China team fully adheres to the safety and security policies of the iGEM competition. All experimental waste in this laboratory is classified according to the level of laboratory waste equipment and recycled by a professional company.
Non pollution level (packaging bags, waste paper, etc.): Put them in black plastic bags and have them cleaned regularly by cleaning personnel.
Medium pollution level (gun heads that come into contact with bacterial strains, waste culture media, etc.): After being sterilized together and ensuring safety, cleaning personnel should regularly clean them.
High pollution level (nucleic acid dye waste glue, etc.): Put it into a collection bag with a yellow biological hazard mark for unified collection, seal it, and then have the recycling personnel of a professional company come to handle it.
Sharp tool level (broken glassware, etc.): Collect in a box with a sharp yellow warning sign, and have the recycling personnel of a professional company handle it at their doorstep.
Non pollution level (water, glucose solution, etc.): Directly discharge after confirming safety and no pollution.
Medium pollution level (culture medium, non-toxic organic solvents, etc.): After classification and sterilization, place them in different waste liquid barrels and have them cleaned regularly by professional personnel.
High pollution level (toxic organic solvents): Put them into a waste liquid bucket with yellow biochemical hazard signs and collect them uniformly (some organic solvents need to be diluted). After sealing, professional company recycling personnel will come to handle them.
