Team:PuiChing Macau/Safety

General lab safety

     The laboratory we use is classified as biosafety Level 1, which shows that the microorganisms we use are almost harmless to human beings. All biological waste will be sterilized with before disposal to make sure that no genetically modified organism is disposed of in the environment.

    In our lab, the safety of researcher are ensured, eye washers/baths and personnel showers in easily reachable positions, installing fire alarms, having fire extinguishers and blankets are provided near the door, having vinyl floors and a ceiling which is inert, and paint on the wall that is easy to clean. There also have view panels on doors to ensure that the risk of accident or injury to researcher whilst entering/leaving the lab is mitigated.

Training and precaution in our team

    Safety and Security training has been given to all students before any experiments. Instructions of experiments will be given to students beforehand. Advice and guidelines from the Faculty of Health Science of the University of Macau are provided. This allow the PI, Ms. Wai Man Cheong, who has been managing a laboratory in our school for decades, to oversee the iGEM team risk management procedures and receive suggestions from experts should it be outside their range of experience. Risk Assessment forms for individual procedures and guidelines for using chemicals were introduced to all students before the project lab experiment start. All experiments will be performed under instructions or supervision of instructors, technicians, professors or teachers (all with appropriate higher education degree).

Rules of our lab

  • No eating or drinking will be allowed in the lab to make sure no students accidentally eat any genetically modified organism.
  • Toe shoes, contact lenses are also prohibiting from entering the laboratory.
  • Liquid biological waste e.g. bacterial culture supernatants will also undergo heat inactivation by autoclaving. The product then can be disposed of via the normal drain.
  • Sharps waste in sharps containers (e.g. needles, scalpel blades) will be disposed of via the clinical waste stream for heat treatment. Spills of biological waste will be cleaned with disposable paper towels and benches will be disinfected with 70% ethanol.
  • Goggles and filter screens are provided to protect users' eyes from intense blue light.

Also, safe use of lab equipment, e.g centrifuge, was demonstrated prior to allowing the use in the lab.

Lab in Macau

    In Macau, there are guidelines from the government regarding biosafety in research labs, which will help us to manage the risks.

Project - specific biosafety

Only disabled laboratory strains of Escherichia coli that are incapable of colonizing the mammalian intestine (Chart et al, 2001) will be used. Disabled strains of E. coli are also not expected to survive in the environment in competition with the wild type strains.

     The DNA constructs transformed to the recipient strains will not offer any particular survival advantage in the wild, or introduce harmful genes, or gene products. The vectors used are not transmissible, limiting the potential for horizontal gene transfer. Constructs may confer antibiotic resistance to strains.

    To avoid the release of resistance genes into the environment, all E.coli strains transformed with such constructs as well as solids and liquids that were in contact with DNA in general and constructs carrying the genetic information for an antibiotic resistance specifically will be inactivated by autoclaving and then disposed.

Prototype

    We choose Synechocystis sp (FACHB-898), unicellular cyanobacteria, for expressing the Lac1326(Yang et al., 2018), and choose Escherichia coli (DH5a), for plasmid construction. All these organisms are not pathogens. They are classified as biosafety Level 1 according to ATCC. They should present minimal potential hazard to the lab and environment.

    The cyanobacteria, Synechocystis sp FACHB-898, and DH5a should restricted to use inside a well-controlled system, such as laboratory. The cyanobacteria and DH5a are non-pathogeni. Also, the laccase is naturally existing to the environment and no report about the serious toxic effect of laccase to animals. Therefore, there is not a large risk for safety, even if unexpected "escape" of the materials to the environment. Even though laccase can potentially influence our immune response and/or chemical reactions in our gut, but it requires a high dosage. So it is safe unless we directly ingest the engineered cyanobacteria or the medium.

References

  • Macao Special Administrative Region Government Health Bureau Public health laboratory service. Clinical Laboratory Biosafety Guidelines(2009), http://www.ssm.gov.mo/pcainfo/doc/2009LabSafetyGuide1_3.pdf