Team:NUDT CHINA/Safety

Designed Protein Degradation Method Based on

Trim21 And Nanobody

Safety

Safety in Project Design

In our project, we use engineered E.coli which is safe for us to work with, which contains the gene of the Trim21 and the antibody of the specific protein. For cells, we use the Hela and HEK293T to complete our project. In addition, we use lentiviral vectors including the TG006 and the pLV. When the Trim21, the antibody and the protein are combined together, the disappearance of the specific protein will be degraded determining by the specificity of the antibody used. And this disappearance will be easily detected by the technique such as the western blotting or Fluorescent labeling. Several concerns of the safety of our project as well as the laboratory are listed below.

1、We choose E.coli as our chassis organism to be genetically modified in our project. Our chassis bacteria have not acquired any characteristics that would enable them to compromise human immune system/other systems or evade detection and destruction by the former or facilitate spread between people/animals, which makes them harmless from both a personal and public health point of view.

2、Our parts are all safe for human, we ensure that none of the genes or parts that we were assembling would act as virulence factors, and that no known pathogens would be involved in our research.

3、All genes were cloned into either pSB1C3, puc57 or pcdna3.1, all of which are non-conjugative, preventing horizontal transfer of our parts. We use LacI as promoter, which guarantees that our bacteria won’t produce large amounts of protein if not induce the expression.