Cell biosafety: Genetically modified organisms (GMOs) are biologically dangerous because they can introduce modified genetic elements into the environment and threaten the safety of other organisms. When assessing GM risk, the following factors should be considered: (1) Potential characteristics that lead to harmful effects. (2) Assess the potential consequences of these harmful effects, and (3) Assess the probability of an individual's potentially harmful effects occurring.
Fig.1 Consider experimental safety and biological protection to ensure that it does not threaten the environment. An anoxic response element was designed (as shown in Fig. 1). when the cell is in anoxic state, an hypoxia-inducible factor (HIF-1α) is produced, which binds to an anoxic response element, thereby activating the minCMV promoter. Therefore, when cells inadvertently escape into the environment, oxygen can disable the cell promoter, thereby avoiding harmful substances to damage the environment.
Safety of Experimental Operations
Fig.2We carefully evaluated the potential risks of the project, including the organisms used, laboratory conditions, laboratory training and waste management. All information is summarized in a security table. We have a Class 1 laboratory space (open bench for non-pathogenic strains) for molecular tests with laboratory equipment, (1) special safety equipment such as emergency showers and eyewashes, as well as fire extinguishers or fire blankets. (Shown in Fig. 2) (2) Waste treatment: treatment of chemicals, biological products and solid waste. (3) Personal protective equipment.
Fig.3To protect laboratory personnel from infections and other hazards, our team members received biosafety education and received systematic training in laboratory. During the course of experiment, we ensure that the laboratory personnel wear safety protective cloting, gloves, and masks throughout the operation. (Shown in Fig. 3, Fig. 4) For infectious lentival vector and modified vectors, team members received systematic training prior to conducting the experiment and conducted experiments under the supervision of trained laboratory personnel (lab technicians and senior graduate students). To prevent the modified gene from being released into the environment, all biological materials, as well as used media and pipettes will be safely sterilized after the experiment. For lentiviral vectors, materials and equipment used will be treated with benzyl dodecyldimethylammonium bromide (C21H38BrN) in order to avoid potential contamination of the environment. (Shown in Fig. 5) Fig.4Fig.5
Safe Use of Reagents
In order to prevent EB from polluting the environment and endangering human health in the experiment. Therefore, an independent area was set up in the experiment to complete the experiment. (Shown in Fig. 6 and Fig. 7)
Fig.6Fig.7
System Security
1. The RAS-RAF pathway is highly expressed in cancer cells. The expression of normal cell is very low.
2. Normal cells do not have anoxic environment and will not activate the killing system.
3. The viral proteins have no target site in the human body and will not arbitrarily cut the protein sequence in the human body; and there are many viral gene sequences in the human body.
4. TIL cells target cancer cells, and reduce infection of normal cells.
5. ALKBH5 inhibits the WNT pathway, which can inhibit the spread and invasion of cancer cells. Normal cells have do not activate the WNT pathway, so this problem is not considered.
6. we are transforming the patient's own til cells to produce no immune response.
7. ALKBH5 system was designed first. If you can't use the killing system, you can prevent excessive killing and avoid the side effects caused by over-treatment.
Lanzhou University
Academic Affair Office of LZU
School of Life Science , LZU
The First Hospital of LZU
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Fig.1 Consider experimental safety and biological protection to ensure that it does not threaten the environment. An anoxic response element was designed (as shown in Fig. 1). when the cell is in anoxic state, an hypoxia-inducible factor (HIF-1α) is produced, which binds to an anoxic response element, thereby activating the minCMV promoter. Therefore, when cells inadvertently escape into the environment, oxygen can disable the cell promoter, thereby avoiding harmful substances to damage the environment.
