We have carefully considered general lab safety, biosafety, animal ethics and laser safety. We choose E. coli. Nissle 1917 and B. subtilis for our molecular cloning and expression. It has been demonstrated that these cells pose no harm to individuals or environment. E. coli DH5α was applied for the experiments of molecular cloning and expression. Some of our experiments are performed on cell lines including MCF 10A, MCF 7 and MDA-MB-231 cell lines. Mouse models are used for the extraction of intended RNA from tumor tissues. They are also needed for the demonstration of the functions of engineered bacteria on tumor tissues. Because mouse models with in situ tumors have great similarities with human tumor microenvironment, they provide a feasible way to assess the antitumor effects of engineered bacteria.

All team members accept responsibility for determining that engineering, testing and measurement involving the use of animals fulfill the international, national and institutional principles, policies, and regulations. Animals sacrificed in this project are highly valued and respected. Research protocols involving the use of animals are in accordance with the national and institutional ethics relevant regulations and guidelines. Any unnecessary discomfort or pain to the animals are avoided. Before the implementation, experimental protocols are carefully designed and strictly assessed whether alternatives exist to animal research. Experiments are performed on cell lines as much as possible. Only in the last steps, mice models are used for the demonstration of the effects of our engineered bacteria. Only personnel who have been trained in animal handling can participate animal experiments.

Our team is committed to maintaining high standards for the lab safety and use of animals. We adopt national and institutional principles for lab safety and the utilization of animals used in testing, research and training programs. All experiments strictly comply with all applicable national, institutional and local laws that regulate the lab safety and animal ethics. All team members accept responsibility for determining the risk factors and fulfill these principles, policies, and regulations. We have provided safety training to team members including general lab safety, biosafety, laser safety and animal ethics.