Tokyo/Genetic circuit

From 2007.igem.org

Abstract   Concept & Model   Requirements   Genetic_circuit   Works   About_our_team

Genetic Circuit

Fig.1: Genetic Circuit
All cells, workers and idlers, have the same genetic circuit. Workers express RFP; while, idlers GFP.

Fig.1 is an outline of the genetic circuit in our model, designed to realize our Concept & Model. All cells, workers and idlers, have THE SAME genetic circuit (just as 'society' members have practically the same genetic backgrounds).

Fig.2: Workers (State A)
A worker, an A-state cell expresses the Pa’s downstream genes including the repressor Ra, LuxI, and RFP. The Pb and its downstream genes for idlers are positively repressed.

Workers (State A)

As shown in Fig. 2, an A-state cell, in which the promoter Pa is "on," expresses the repressor Ra, LuxI, and RFP. The repressor Ra "turns off" the promoter Pb and inhibits the cell-state change. This cell also expresses LuxI, which makes AHL, cell-cell communication molecules. Downstream of these genes, there is an RFP gene; therefore, we would be able to detect A-state cell using flow cytometer.

Idlers (State B)

Fig. 3 shows a B-state cell, in which promoter Pb is "turned on" in the presence of AHL, and repressor Rb and GFP are expressed. Repressor Rb represses the expression of promoter Pa and thereby RFP; therefore, fluorescence of GFP is used to detect B-state cells.

Fig.3: Idlers (State B)
In the presence of enough AHL, an idler or a B-state cell expresses the Pb’s downstream genes including the repressor Rb and GFP. The Pa and its downstream genes for workers are positively repressed.

If Workers are removed...

As shown in Fig 4, if A-state cells are removed, B-state cells sense the absence of A-state cells. Responding to the decrease of AHL, cells stop producing repressor Rb. Now that repression of Pa by Rb as well as that of Pb by Ra are declined, either state cannot be clearly expressed, as if the cells were reset.

Fig.4: If Workers are removed...
If workers are removed, AHL concentration decreases, which declines activation of promoter Pb. With less gene expression downstream of Pb, the B-state becomes unstable.

Hybrid promoter

To realize such a genetic circuit, Pb should be "Hybrid promoter" regulated by AHL and LacI as shown in the figures.

Other ideas of genetic circuit

Also, we thought of other ideas of genetic circuits which perform the same function.

The ideas are here.