Team:UT-Tokyo/Description
Description
We aim to create a genetic circuit functional in E.coli which allows the gene expressed to change after each cell division. Specifically, the E. coli will be expressing GFP, RFP, and CFP after subsequent cell divisions and will loop back to expressing GFP again.
This will be achieved by the use of the Pnrd promoter to sense to cell division, sigma factor/anti-sigma pairs control transcription, and a toehold switch to create an AND gate required in the circuit.
Our project would be able to exhibit, in E. coli, that despite carrying the same genetic information, different phenotypes can be expressed across generations. Much like the phenomenon seen in epigenetics. Moreover, this technology could potentially be applied in field to carry out certain tasks automatically without the need of external input eg. to make engineered cells automatically carry programmed cell death after a certain number of divisions.
Parts Improvement
Stanford-Brown iGEM 2012 documented the promoter Pnrd (BBa_K847211 ) which activates during cell division and confirmed its functionality. However, since it was submitted in RFC[23] standard, we decided to improve the part by using redesigning the part in RFC[10 ] (BBa_ K2070012) in order to deal with the disadvantages of RFC[23 ] standard. Details on the RFC[10 ] and RFC[23 ] standards can be found on the iGEM wiki.
