"
Team:Evry/AIDSystem
From 2012.igem.org
Orthogonal hormonal system
We adapted the auxin production device from the iGEM team Imperial college 2007 to eucaryotes and combined it with an auxin detection module. This way, we created the first synthetic hormonal system for intertissues communication.
To test this system, we co-injected plasmids expressing our production and reception devices in tadpole's embryos, a new chassis we wanted to implement for synthetic biology. We performed auxin toxicity and uptake tests at the begining of our project to ensure the feasability.
Auxin production devices
We envisaged 3 ways of design our auxin production device in tadpoles. The devices 1 and 2 have been designed to be expressed in tadpole's embryos, while device 3 have been designed to be expressed in E.coli. In this last case, the aim is that the tadpoles eat bacteria expressing device 3.
- Auxin production device 1 : this device is composed of BBa_K812021, coding for IaaM, and BBa_K812120, coding for IaaH for auxin generator for the use in tadpole's embryos.
- Auxin production device 2 : this device is composed of BBa_K812014. It is ment for the co-expression of IaaH and IaaM genes in the same cell in tadpole.
- Auxin production device 3 : this device is composed of BBa_K515100, coding for IaaM and IaaH for auxin generator in E.coli.
Pathway
Auxin reception devices
We envisaged 2 ways of design our auxin production device in tadpoles. To visualize the communication between different tissues or between E.coli and a tissue of the tadpole, we chose to work with GFP.Our orthogonal hormonal system works with any proteins fused to AID signal with any transcription factors.
- Auxin reception device 1 : this device is composed of BBa_K812010 coding for GFP-AID, and BBa_K812012 coding for OsTir1.
- Auxin reception device 2 : this device is composed of BBa_K812013 coding for GFP-AID and OsTir1 in the same cell.
References:
- 1. Nishimura, K., Fukagawa, T., Takisawa, H., Kakimoto, T. & Kanemaki, M. An auxin-based degron system for the rapid depletion of proteins in nonplant cells. Nature methods 6, 917-22 (2009).
