Difference between revisions of "Part:BBa K3365065"

Revision as of 10:52, 25 October 2020

Fourth lure sequence upstream of eGFP

This part is a off-target detecting platform for dCas9-ω. We put this part in a plasmid and transfer it into △rpoZ-MG1655, a strain of E.coli that was koncked out rpoZ. If dCas9-ω is binding to this lure sequence, the submit Omega fused to dCas9 C-terminal can activate the expression of reporter by recruiting the RNAP holoenzyme and the cell would emit green fluorescence,which means dCas9-ωis off-target, else the weak promoter and the lack of rpoZ makes the reporter cannot express or just have a quite low expression level.

According to the prediction model, the off-target rate of the lure sequence in this part is 0.37543%.

Fig:Schematic diagram of this part

Sequence and Features