Since its development nearly a decade ago, DNA origami has served as a powerful tool for the creation of self-assembling functional materials (such as proteins) on a nanoscale in both 2- and 3-dimensions. Unfortunately, the complex formations built using a DNA origami template lose their created structure upon any damage to or removal of the DNA scaffold. There is a pressing need to demonstrate that the structural organization of these functional materials can be maintained after separation from the DNA origami template upon which they were built. This work attempted to merge the unique structural possibilities of DNA origami with the biological relevance of proteins in order to achieve ordered protein constructions on a DNA origami scaffold; chemical cross-linking was used to fix the proteins into position so that they would retain their shape upon the destruction of the scaffold.<br/><br/>Two polyethylene glycol (PEG) based protein cross-linkers of different lengths (MW: 2000 and 6000) were synthesized. Rectangular DNA origami pieces were prepared in accordance with previous literature. In this case, however, several of the staple DNA strands for the structure had been biotinylated in order to have programmed biotin locations on the template. Streptavidin (StAv) was later immobilized onto the origami surface through binding with the introduced biotin moieties, followed by incubation with either of the two PEG cross-linkers. Crosslinked StAv array was obtained after the destruction of the DNA origami structure and further purification. Atomic Force Microscopy (AFM) was used to characterize each sample through the steps of this process. AFM results showed that the cross-linked StAv retained its designed arrangement after the scaffold had been removed.

Summer research in Ulm, Germany: Ordered protein assembly via a DNA origami scaffold

251st National Meeting (2016)

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