Team:SCUT-China/Safety
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SAFETY
Laboratory Safety
In our project, we used BL1 materials E. coli for cloning and BL2 materials Human Embryonic Kidney 293 cells for transfection. All students in our team are well-trained. Students who worked with BL2 material received blood-borne pathogens training besides usual lab training. Before our experiment, we made a detailed protocol and communicated with our instructor to ensure that we met the criterion in laboratory. When doing experiment, we wore lab coat, gloves and gauze mask to protect ourselves and all the experiments of BL2 materials were handled in biosafety cabinets. In accordance with MSDS, after finishing experiment, we handled carefully all laboratory chemicals, especially BL2 materials, which were handled with decontamination bleach mixture under ultraviolet rays for 24 hours.
Biological Parts
We constructed 2 basic parts that encoding internal protein in human body and designed 3 scilencing devices . The most hazardous biological parts in our parts are the scilencing devices since they encode the hairpin RNA that do not exsist in human body and they may cause potential risks. We transfected the scilencing devices into HEK293 to scilence the PDE5A gene. To make our parts safer, we designed a hypoxia responsive switch, only under hypoxia situation the down stream parts will work. In our experiment we used lentivirus to packaged our plasmids so that all the transfection experiments were handled in II biological safety cabinet.
Bio-safety of lentivirus vectors
The lentiviral vectors we used in our project are self-inactivating virus, which means that after infecting target cells， virus will not infect other cells, and will not produce new virus.Although we minimized the risk by modifying the vector design, insertional mutagenesis caused by lentiverus still cause unknowm risks. Quantitative assays have demonstrated that the risk of insertional up regulation of proto-oncogenes is directly related to the strength of the enhancer sequences contained in the vector. Thus before we use lentivirus vectors for clinical purpose, a series of cells and animals experiment should be done to evaluated the safety of lentivirus vectors.