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An issue with a previous model (DNA Origami Glue) was that in order to create complex unique shapes with specifically bonded cells at a chosen location there needed to be enormous amounts of unique zinc-fingers.
The aim of this model is to design a 3D, self-assembling structure which forms a shape which allows E-coli cells to be bonded to the outside. One of the shapes we decided to use is a geodesic sphere made up of multiple tetrahedron ‘bricks’. Tetrahedrons are the strongest 3D structure and would allow any sized scaffold to be made. A sphere is the best 3D structure as it has the largest surface area to volume ratio which will allow the largest number of E.coli cells to bond to it.
The first two images are how the tetrahedrons are made, using DNA origami. Our sequences will be designed so that each side will have DNA hairpins which bond to another side of a different tetrahedron. The last image is what the DNA origami structure will look like once it has been made.
Each side of the tetrahedrons will have the binding sequences needed to bind the E.coli cells to the outside of the structure, so that it doesn’t matter where each tetrahedron goes.
These images show how each side of the tetrahedron will have different hairpins. The red side will bond to the yellow side, and as you can see they can form many shapes.
Optimising the size of the tetrahedrons
The smaller the tetrahedrons the stronger they are and hence the stronger the resulting formed 3D structure will be. However by decreasing their size you increase the amount of DNA you need to construct it which adds complexity, takes more time and is more expensive. Therefore it is important to find a compromise between size and amount of DNA used.
From reading various papers, such as this one we determined the maximum size you could make was a tetrahedron with side lengths of 75nm. This size maximised stiffness and strength while minimising the amount of DNA.
This graph shows the exponential increase of the number of tetrahedrons needed to bond various amounts of E.coli cells to the surface of the formed structure. Due to our time period and budget it would seem a lot better to use a smaller value of E.coli cells to be bonded (<50) to minimise the DNA sequencing.
