Minimizing the risks for team members and society is one of the most important issues, especially for us undergraduates. We shall further illustrate the concept of biosafety and our endeavors by quoting and explaining the five guiding principles of safe manipulation gene manipulation biology (GMO) summarized by Kimman et al. in 2008[1].
1. Hazard identification: Identification of risk assessment have been completed, and all laws and regulations have been considered in accordance with the provisions of the last paragraph.
2. Biological containment: Biological containment refers to the use of organisms with "reduced replication capacity, ineffectiveness, transmissibility and toxicity". For our project, replication-restricted viruses are used and they are all self-inactiviting lentivirus.
3. Concentration and Shell: All working steps for packaging lentivirus are carried out in our BSL II laboratory, and lentivirus is limited to a biosafety cabinet. Cell culture and virus storage are also carried out in separate incubators and freezers.
4. Exposure minimization: This aspect of the exposure minimization guidelines can be performed under "Operator Protection". When the manipulating the viral vectors, care should be taken to avoid liquid droplets, especially aerosols. By inserting or removing materials from laminar flow cabinet and cleaning all equipment after completing the work tasks, we can avoid the possible transfer of virus carriers.
5. Physical containment: We will meet the requirements of physical containment by performing all operations on lentivirus in BSL II laboratory in which we shall guarantee the number restriction for persons’ entrance in the BSL II laboratory.

Managing the risks

Rules and guidance

The General requirements for laboratory biosafety are the overall rules of biosafety, keeping most of our actions in the laboratory safe. And most norms in these requirements comply with international standards. In this handbook, Laboratories—General Requirements for biosafety, enacted by the People’s Republic of China (PRC), stated the security classifications, laboratory managements, precautious strategies, waste disposal and all kinds of general requirements and regulations an researcher might encounter in conducting biological experiments. Below is the link of this handbook. GB 19489-2008 Laboratories-General requirements for biosafety.

Laboratory training

All our participants have received relevant training on biosafety, and received laboratory standard operation and ethics training in the course. We have promised to pay much attention to physical protection on our body and strictly follow the standard procedures and protocols provided by team advisors and company. Before entering the laboratory, adequate disinfection and change of clothes, shoes, masks, hats and gloves are required.

Safety of chemicals

The main toxic and harmful chemicals uesd in our project are ethidium bromide(EB), trichloromethane(CHCl3) and Trizol. To prevent EB used in agarose gel electrophoresis from polluting the laboratory, an independent region was set in the labotatory to use EB. And we use trichloromethane and Trizol in a chemical fume hood when we extract RNA.

Waste disposal

Additionally, the purchase, collection, storage, use, transfer and waste disposal of hazardous chemicals must be strictly implemented in accordance with national laws and regulations and the relevant provisions of the school. Biosafety cabinets and instrument materials are fully sterilized before and after use, including alcohol&benzalkonium bromide treatment and UV exposure.

Discussion about ethic in the use of RNA editing

Compared with DNA editing like CRISPR-Cas9, RNA editing doesn’t cause permanent changes to the genome, and this reversible and easily regulated editing method may have security advantages. In the 2021 CCiC meetup, Zhao GuoPing, who is an academician of Chinese Academy of Sciences, also mentioned that RNA editing doesn’t face the same ethical challenges as Cas9.


[1] Kimman, Tjeerd G., Eric Smit, and Michel R. Klein. "Evidence-Based Biosafety: a Review of the Principles and Effectiveness of Microbiological Containment Measures." Clinical Microbiology Reviews 21.3 (2008): 403-425.