Team:Warwick/Cloning
Cloning and Transformation
Our project relied greatly on previously established techniques for cloning modified genes into plasmids and transforming those plasmids into competent cells. We wanted to make our part as modular and easy to change as possible, meaning we had to rely heavily on techniques such as Gibson assembly to construct our plasmid. By offering our part with changeable parts we hoped to make our part more useful for further research, and allow for deeper applications than the ones we initially thought of.
Having interchangeable zinc fingers allows the diversity of the cells that can be glued together using our system to be much greater by allowing each different cell type or individual cell to have a specific DNA sequence that it binds to, so that fine control of spatial positioning of cells can be achieved.
By having modular anchor domains, we allow for the expression of the zinc finger on the surface of the cell to be controlled as well. By changing the anchor domain, you affect the number of zinc fingers expressed on the surface of the cell, as well as their binding affinity, due to different anchors forcing the zinc fingers closer or further away from the cell. Our design also includes unique cut sites for the linkers in the part, so that they too can be cut out and changed for longer or shorter ones, also affecting zinc finger expression or binding.
This diagram shows our starting point in the project with the JO4450 biobrick part expressing RFP. It also shows the modularity in our design with the interchangeable anchor domains and zinc finger domains.