Team:Tsinghua/Safety

Yeasy AFT project page

Overview

This year, Team Tsinghua proposed a yeast-two-hybrid-based household aflatoxin detection system and attempted to realize such design in Saccharomyces cerevisiae, of which the biosafety level is well recognized and the protocol for experimental manipulation is well established. Although AFT we used is cancerogenic, we made sure that every step is under rigid control so that no hazardous chemical will be let out. We consulted the PI from labs that have been studying aflatoxin for a relatively long time for safety protocol when working with it.
Safe though it may seem, there still could have been unsolicited consequences if no attention for safe experiments is heeded. In order to minimize any unwanted accident, we received proper training and adopted several safety regulations in the laboratory. Now we’d like to share our experiences with safety control.

Safe project design

Firstly, we choose a non-pathogenic chassis for our project. We employ Saccharomyces cerevisiae as our chassis and E. coli for the molecular clone. All the strains of yeast and E. coli we used are auxotrophy and laboratory waste must go through the sterilization process before released to the environment to prevent the escape of engineered microorganism. Secondly, as the aim of our project is to develop a bio-system that detects the presence of aflatoxin and gives a warning for that, we engineered the sequence of aflatoxin antibody into yeast, fused to the yeast two-hybrid elements. This protein contains a fragment of the antibody, but not a fragment of aflatoxin itself. The usage of aflatoxin is limited in a few steps of the whole project. Thirdly, to reduce the risk of aflatoxin use to researchers, we used productization aflatoxin only from certified suppliers, and only buy and use aflatoxin solutions rather than dry powders, which avoids inhaling of the powder. We also took strict measures of experimental operation to protect researchers and prevent aflatoxin contamination.

Safe lab work

Before we carried out our experiments, all of our team members had received strict lab safety training, including standard experimental protocols, proper disposal of biological and chemical waste, information on hazardous chemicals and use of biosafety cabinets given by PI in the lab.
Such guidance was documented on the online portal in detail. Team members involved in bench works had all went through orientations with mentors or managers in charge of specific equipment used before gaining access to the lab.
According to the requirements of iGEM policy, we never performed any dangerous experiments in daily bench work or faced any unusual safety issues. We followed some rigid rules while carrying out all the bench works:

  • 1. All participants are supposed to pass a test including principles and operations of the experiment before commencing work.
  • 2. All experiments are done with rubber gloves.
  • 3. Always use appropriate (BSL-2 standard) personal protection equipment (lab coat, gloves, goggles, mask) when working with aflatoxin.
  • 4. All experiments involving the use of aflatoxin or having the potential of live microorganism release are done in biosafety cabinets.
  • 5. Hazardous chemicals such as ethidium bromide (EB) and AFT were carefully used and disposed of according to protocols.
  • 6. No food or drink is allowed in the working area.
  • 7. All appliances, especially microwaves and centrifugal machines are supposed to be used with full-time attendance.
  • 8. All wastes should go through a rigid disposal process before discarding.
  • 9. The last person to leave the lab should always make sure to shut down all electricity and water supply before leaving.

We especially focused on the safety of AFT-related experiments. Everyone involved in these experiments was specifically trained and all the steps are carried out in high-security labs. With the help of other labs, we established a specific process to dispose of AFT after experiments. We expect to minimize the exposure of AFT to human body and environment.

Overview

All DNA parts were transported and operated following Parts Registry requires.

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