Design and synthesis of DNA origami nanostructures to control TNF receptor activation

Clustering of type II tumour necrosis factor (TNF) receptors (TNFRs) is essential for their activation, yet currently available drugs fail to activate signalling. Some strategies aim to cluster TNFR by using multivalent streptavidin or scaffolds based on dextran or graphene. However, these strategies do not allow for control of the valency or spatial organisation of the ligands, and consequently control of the TNFR activation is not optimal. DNA origami nanostructures allow nanometre-precise control of the spatial organization of molecules and complexes, with defined spacing, number and valency. Here, we demonstrate the design and characterisation of a DNA origami nanostructure that can be decorated with an engineered single-chain TNF-related apoptosis-inducing ligand (SC-TRAIL) complexes, which show increased cell killing compared to SC-TRAIL alone on Jurkat cells. The information in this chapter can be used as a basis to decorate DNA origami nanostructures with various proteins, complexes or other biomolecules.