Professor Andrew Turberfield

Abstract:

Many DNA nanostructures have been produced and a wide range of potential applications have been proposed. However, confirmation of accurate 3D construction is particularly challenging. Here, we demonstrate that cryoEM may be exploited to obtain structural information at sufficient resolution to visualize the DNA helix and reveal the absolute stereochemistry of a 7 nm self-assembled DNA tetrahedron. Structural analysis at such high resolution by cryoEM image analysis is unprecedented for any biological molecule of this size.

Abstract:

We present a facile method for linking recombinant proteins to DNA. It is based on the nickel-mediated interaction between a hexahistidine tag (His(6)-tag) and DNA functionalized with three nitrilotriacetic acid (NTA) groups. The resulting DNA-protein linkage is site-specific. It can be broken quickly and controllably by the addition of a chelating agent that binds nickel. We have used this new linker to bind proteins to a variety of DNA motifs commonly used in the fabrication of nanostructures by DNA self-assembly.

Abstract:

Metastable two-stranded DNA loops can be assembled into extended DNA oligomers by kinetically controlled self-assembly. Along the designed reaction pathway, the sequence of hybridization reactions is controlled by progressively revealing toeholds required to initiate strand-displacement reactions. The product length depends inversely on seed concentration and ranges from a few hundred to several thousand base-pairs.