Team:BostonU/Temporal Control/Current Methods
| Overview | Current Methods | Our Workflow |
Current Methods
One way to temporally control a protein is to physically split it and fuse halves of the proteins to domains that naturally dimerize in the presence of an inducer; and absence or presence of inducer will modulate when the protein is inactive or active, respectively.
| Another way to control proteins is to put it under the control of an inducible promoter. This is a type of transcriptional control. Transcriptional control is an effective method of creating single logic circuits in cells. | INSERT SIMPLE AND/OR/NOR GATE WITH INDUCIBLE PROMOTER LOGIC GATE |
SHOW IMAGE OF HIGHER ORDER INDUCIBLE PROMOTER LOGIC GATE |
However, building nested circuits and higher order logic becomes challenging with this system. |
The nested logic causes delays in response with each layer of logic. In post-translational modulation systems, the proteins are fully functional as a response to the proper inputs.
For the split integrase + RDF and saCas9 systems that we explored, it was important that for the input control to be completely orthogonal to the cellular system. The integrase + RDF system would not supply any output with inducible promoters that allowed transcription before an input signal was received. In other words, any leakiness at all in the inducible promoter system with integrases and RDFs would cause transcription and translation and turn the gene immediately off. Post-translational control by splitting gives us tighter control over the integrase and RDF system.