SAFE DESIGN
All DNA sequences used for constructing the biosensor were ordered from company IDT. They are T7 promoter, binding site sequence for transcription factor ArsR, fluoride riboswitch sequence from Bacillus cereus, iSpinach aptamer and its F30 scaffold, bubble duplex and Anderson promoter. The only protein sequence we used is sfGFP, that was cloned from pJUMP-48-2A plasmid using PCR. They are not pathogenic and harmful in anyways to human and environment.
Our system is cell-free, which uses cell extract only. Thus, no GMO issue. Cell extract we used were from E. coli K12 MG1655 and Cupriavidus metallidurans CH34, which are known to have biosafety level 1.
The coolest thing about our project is the fact that it is cell-free! We harness the mechanism of transcription of a cell without the need to keep it alive. This gives us better control over the transcription mechanisms, without the problem of metabolic burden, reproduction, and cell wall permeability.
