Project safety has always been a very important part of biological research, especially in iGEM. All experiments in our project are based on the promise of safety. In order to reduce the safety risk of the laboratory, team members have received safety training before entering the laboratory. Each team members attaches great importance to the safety requirements of laboratory work. Throughout the project, team members followed the iGEM safety guidelines strictly.
Safety in lab
According to safety protocols, our lab is classified as Biosafety level 1. We received safety training before starting the experiment, learning and following safety protocols, and took care to ensure safety during the experiment.
White lab coat, mask and rubber gloves are required for laboratory work;
·Before starting the experiment, read the instructions carefully and be familiar with the operation of the equipment to avoid the accidents caused by the unfamiliar operation;
·For the use of sterilization pot we strictly follow the standard process;
·Most of the reagents used in this experiment are safe;
·Wear rubber gloves when using reagents to avoid skin contamination with toxic reagent (such as DMSO);
·Standardize experimental operation to avoid operational errors;
·The laboratory separates all the experimental materials from dry and wet;
·The bacteria liquid used in the laboratory is collected, processed and then discarded, which reduces the possibility of laboratory bacteria contamination of the environment.
We also have lab directors who manage our safety affairs, and we have institutions at the college and school level that act as supervisors and supervisors of lab safety.
Safety in project
We try to use the CRISPR-Cas9 system to cut all of the Saccharomyces cerevisiae's chromosomes. As cellular machinery remains functional in these chromosome-free cells, they can be used to complete the delivery of large fragments of DNA or macromolecule drugs.
Firstly, as our organisms lose all of its' chromosomes, they cannot reproduce and grow autonomously in theory. Besides, they have a short life cycle which means they will automatically die. These features are very effective in ensuring its safety and reduce the possibility of harm to the environment. Secondly, in this experiment, we used Escherichia coli as the chassis for molecular cloning and Saccharomyces cerevisiae as the modified strain. Both strains are widely known and non-pathogenic. Thirdly, our strains are auxotrophic. They must be added with specific amino acids such as histidine, leucine, and uracil in order to grow. Fourthly, the bio-parts used in this experiment are derived from safe plasmids, so they are not unknown segments, reducing the possibility of environmental pollution, pathogenicity and allergies.
In summary, this project can effectively prevent bacteria from escaping from the laboratory and complete the cell apoptosis process to ensure the safety of the entire project and avoid environmental pollution.
Safety in human practice
Although our project has high safety, since our goal is to eventually apply CREATE to drug delivery in human body, through our questionnaire survey, people generally have higher safety requirements for microbes used in human body. To this end, we interviewed Professor Song Xinyu from Tianjin University Center for Biosafety Research and Strategy which is the only Biosafety Center in China.
Professor Song xinyu introduced us to the Tianjin Biosecurity Guidelines for Codes of Conduct for Scientists and answered questions about our project. As for the direction of drug delivery envisaged by us, Based on her own research and the content of the paper, Professor Song Xinyu concluded that the potential risks of drug delivery could be divided into the following two parts: biosafety and bioethics.
For our project, Professor Song Xinyu also gave professional suggestions. From the experimental point of view, because there is no chromosome, it is difficult to replicate, indeed to a certain extent to ensure biological safety. However, if it is applied to drug delivery, it is impossible to guarantee whether other organelles, cell membranes and other structures of yeast will cause allergies, and in the process of drug delivery, it is difficult to guarantee that there is no miss target problem.
From the perspective of ethics, since it is impossible to define whether it belongs to a kind of organism, it is difficult to determine whether it belongs to genetic engineering products or exogenous organisms, and whether it can be put into the market needs subsequent bioethical review. Moreover, drug delivery needs to consider more biosafety issues, not only the risk itself, but also the patient's informed consent right. Therefore, in the subsequent development of downstream technology, CREAT cells still have many parts worthy of in-depth discussion.
Under the advice of the professor, we will make two preparations next: we will ask the scientific researchers about the significance of the project and get research guidance and suggestions; Popularizing biological engineering to the public in order that it can be recognized by the public.
The professor also fully appreciated our project. She believed that with the popularization of bioengineering, more and more people will know bioengineering, bioengineering management will be more and more standardized, biosafety will be more and more valued, and people will accept biological products more and more. At the same time, however, she expressed concern about the increasing risk of misuse of the technology, with biosafety becoming an increasingly serious issue as bioengineering becomes more widespread.