Safety


Overview

Squirrel-Shenzhen recognizes that biosafety and biosecurity are critical to the success of the entire iGEM project. Therefore, we have divided safety into two parts: laboratory safety and biosafety. For laboratory safety, we have ensured the biosafety of the chassis organisms and parts, as well as confirmed the proper disposal of laboratory waste liquids containing copper and zinc elements. Additionally, under the guidance of our instructor, we completed two laboratory safety training sessions and followed laboratory management protocols for lab operations and waste management. For biosafety, we have chosen a light-controlled suicide system as our biosafety mechanism to prevent bacterial and genetic leakage. We understand the inherent risks of working in laboratory facilities and are committed to taking all necessary precautions to prevent harm to individuals and the environment.


Lab Safety
Chassis and Part

The chassis we used are E.coli DH5a and E.coli Mach1-T1, which all belong to RISK GROUP 1, means they are low risk for human being and environment.

Our parts are all taken from the RISK GROUP 1 and RISK GROUP 2 organisms. The RISK GROUP 1 organisms are not safe to cause diseases in healthy adults. Although the RISK GROUP 2 organisms may cause diseases in humans, the parts we extracted were all synthesized from the company, without any risk of pathogens.


Disposal

In our laboratory, we have designated containers for solid waste, waste liquids, and sharps, all of which are managed and collected by Jet Biofil. We are required to fill out labels, clearly indicating the "date, lab location, and type of waste," and attach them to the waste bags or liquid waste containers for centralized collection and disposal by Jet Biofil.

Additionally, to prevent environmental contamination, we collect all bacterial culture media and sterilize them before disposal. To maintain laboratory safety and cleanliness, we conduct regular cleaning, sterilization, and disinfection on a daily basis. The clean bench is sterilized with UV light before use.


Training

Our team members participated in two laboratory safety training sessions.

The first training was an online course conducted by our Secondary PI, Riva Yan, who explained laboratory safety, including the Safety Form, daily laboratory management regulations, safe usage of water, electricity, fire, and gas in the laboratory, safe handling of chemical reagents, hazards and management of microorganisms, safe usage of radioactive materials, the hazards and protection against UV radiation, and waste management.

The second training was an in-person session held in the laboratory, where our instructor, Rong, provided hands-on safety instruction, covering the use and precautions of basic laboratory equipment, the operation and precautions of the clean bench, waste management, daily laboratory hygiene and safety management, and the use of emergency escape routes.


Operation

Our laboratory operations strictly adhere to the safety regulations of the (PDF file below). Additionally, we have established the following daily laboratory rules:

  1. Lab coats must be worn at all times while working in the laboratory.
  2. When handling volatile toxic reagents, operations must be conducted in a fume hood.
  3. All reagents have designated storage locations and must be returned after use.
  4. All chemicals and reagents must have clean, clearly marked labels, including the name, concentration, and specifications.
  5. After completing work, all laboratory surfaces must be disinfected daily.
  6. Eating, drinking, and smoking are prohibited in the laboratory.
  7. Mouth pipetting is strictly forbidden. Lab-provided pipettes, syringes, and pipettors must always be used.
  8. Contaminated glassware, plasticware, and discarded petri dishes must be placed in containers designated by the supervising coach.

Overview



Biosafety

To prevent bacterial leakage, we have implemented strict engineered eradication measures—a light sterilization system. We selected a light-activated suicide switch, ensuring that E. coli can be totally killed in UV. This system consists of a UV light-sensitive promoter switch (pUV l765001 - BBa_K4186000) and the lysin gene Lysep3-D8 (BBa_K3034004). The lysin gene is a composite protein that can lyse bacteria both inside and outside the cell, theoretically exhibiting stronger bacterial lysis effects. We have completed both the plasmid and experimental design, but due to time constraints, the experimental validation has not yet been completed.

The three plasmid diagrams in represent the suicide gene system and its control experiments. To verify the function of the lysozyme and the light-inducible promoter, we designed three plasmid combinations: pUV l765001+Lysep3-D8, pUV l765001 + dsRed, Tet + Lysep3-D8, which are located in the upper left, upper right, and lower sections.

Safety Form

Final Safety Form: Link to Safety Form