Team:SCU-WestChina/safety

Safety

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Safety

The general public has many misconceptions when it comes to genetically modified organisms (GMOs), which then leads to a lot of fear and stigma. In terms of gaining widespread acceptance for synthetic biology applications in daily life, there is still much effort to be made. Since the advent of synthetic biology, scientists have been working hard to implement safeguards in their genetically modified organisms. When it comes to biotherapeutics or any product of genetic engineering that will be consumed by humans, safety has never been more important.

Laboratory Safety

The iGEM SCU_WestChina team is in full compliance with the safety and security policies of the iGEM competition.

The team was included in the laboratories of Prof. Xie Huiqi and Prof. Yang Hao, which contains equipment for both Biosafety Level 1 and 2. efore we started our work in the lab, all team members had to receive training and pass different safety tests provided by our university. We also received training from our advisors on how to use the machinery in the lab (i.e. thermocycler, Surface Plasmon Resonance Machine) and also on the use of chemicals (i.e. p-Cresol, staining reagents). At all times, a supervisor or instructor was present when work was conducted in the laboratory.

Project Safety

We plan to use E.Coli in submerged culture which will be inactivated after our extractin proteins, and we plan to use HFL-1 cell culture in solid culture which will be properly cleaned after our experiments finished.

E. coli BL21(DE3)

In order to make our biotherapeutic safe for human consumption, we used E. coli BL21(DE3) as our chassis. The BL21(DE3) as our The strain contains DE3, a λ prophage carrying the T7 RNA polymerase gene under control of the lac UV5 promoter and lacIq. IPTG is required to induce expression of the T7 RNA polymerase. Protein expression from transforming plasmids containing a T7 promoter-driven expression system is repressed until IPTG induction of T7 RNA polymerase expression occurs. The strain does not contain the lon protease and is also deficient in the outer membrane protease, OmpT. The lack of two key proteases reduces the degradation of heterologous proteinsexpressed in the strain. In this way, the foreign protein expression can be controlled well. And we only apply this strain for protein expression , so it will be inactivated after the protein is extracted.

Our bacteria will be engineered to produce recombinant proteins( combining the probes with receptors ) which will be loaded by liposomes later. They will be applied to digest collagen fibrosis as well as to block key pathway of IPF of human lung fibroblast cells in vitro.

HFL-1

As the European Collection of Authenticated Cell Cultures (ECACC) rules, we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines. ACDP = Advisory Committee on Dangerous Pathogens (UK). The cells were only induced by TGF-β1 to set the cell model for drug efficiency verification, and we plan to use HFL-1 cell culture in solid culture which will be properly cleaned after our experiments finished.

Chemicals

As we usedacrylamide(Acr),methylene bisacrylamide(Bis),APS,TEMED for SDS-PAGE, so we add these chemicals in the hume hood under supervision.

Experts’ Overseeing

The lab technician Zhang Yanjing will ensure the safety and security of biology labs at our institution. She will supervise and guide us in reviewing our work. She is really good at protein expression and purification, SPR, cell culture and other experiments, and she also knows safety and security managements in biology labs well.

Firstly, we will use standarized protocols to finish our experiments with her guidance.

Secondly, after our verification of primary work in vitro during 2020 iGEM, we will apply for later animal studies and clinical trials after getting permission of strict ethical approval. We believe our project will be assessed in various dimensions to provide more efficient therapy for patients.

Guide Rules

1.Compilation of laws and regulations related to biosafety of the People's Republic of China.
This reporting system sorts out the current and effective legal documents related to biosafety in China, including pathogenic microorganisms, high-level biosafety laboratories, infectious disease prevention and control, hospital control, medical waste, emergency Public health incidents, food safety, agricultural genetic modification, biological products, traffic quarantine, animal biosafety, plant biosafety, entry and exit quarantine, protection of genetic resources, dual-use items and technology control, and biotechnology supervision were classified. It can provide reference for people engaged in biosafety-related work in my country.

2.List of pathogenic microorganisms transmitted from humans, China CDC

http://www.nhc.gov.cn/wjw/s9502/201212/34016.shtml

3. All of our wet lab members have been well trained, and we won't use any pathogenic strains of an organism or take up any unwarranted animal experiments. And our waste produced in our project will be successfully inactivated by professional lab mananger in our lab.

Reference

[1] Studier and Moffatt, J. Mol. Biol. 189 (1986), 113-130 PMID: 3537305
[2] Breul SD, et al. Control of collagen production by human diploid lung fibroblasts. J. Biol. Chem. 255: 5250-5260, 1980. PubMed: 7372635

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