Estimation of microtubule-generated forces using a DNA origami nanospring

Microtubules are dynamic cytoskeletal filaments that can generate forces when polymerizing and depolymerizing. Proteins that follow growing or shortening microtubule ends and couple forces to cargo movement are important for a wide range of cellular processes. Quantifying these forces and the composition of protein complexes at dynamic...

Orientation-Locked DNA Origami for Stable Trapping of Small Proteins in the Nanopore Electro-Osmotic Trap

Nanopores are versatile single-molecule sensors offering a simple label-free readout with great sensitivity. We recently introduced the nanopore electro-osmotic trap (NEOtrap) which can trap and sense single unmodified proteins for long times. The trapping is achieved by the electro-osmotic flow (EOF) generated from a DNA-origami sphere...

Towards a bottom-up reconstitution of the nuclear pore complex

At first glance nanopores may appear simple, almost intuitive, to understand given that they are, quite literally, ‘just’ very small pores in a membrane. In fact, one may even wonder why at all we need trained scientists to study such seemingly simple entities. The short answer is that nanopores, as the word suggests, are nanoscale entities and,...

Reconstitution of Ultrawide DNA Origami Pores in Liposomes for Transmembrane Transport of Macromolecules

Molecular traffic across lipid membranes is a vital process in cell biology that involves specialized biological pores with a great variety of pore diameters, from fractions of a nanometer to >30 nm. Creating artificial membrane pores covering similar size and complexity will aid the understanding of transmembrane molecular transport in...

High-Speed Super-Resolution Imaging Using Protein-Assisted DNA-PAINT

Super-resolution imaging allows for the visualization of cellular structures on a nanoscale level. DNA-PAINT (DNA point accumulation in nanoscale topology) is a super-resolution method that depends on the binding and unbinding of DNA imager strands. The current DNA-PAINT technique suffers from slow acquisition due to the low binding rate of...

Plasmonic nanopores for single molecule sensing

Grabbing a single molecule and inspecting its contents is far from easy. Apart from the small size of the objects, biomolecules shake, shimmer, and bounce around a tremendous amount. How can one gently control something that small (without squashing or destroying it) and still be able to tell what it is? The work in this thesis is exactly aimed...

Mimicking the nuclear pore complex using nanopores

Nuclear pore complexes acts as a gatekeeper for molecular transport between the nucleus and the cytoplasm in eukaryotic cells. The central NPC channel is filled with intrinsically disordered FG domains (phenylalanine (F), glycine (G)) that are responsible for the fascinating selectivity of NPCs, for which the underlying mechanism is still under...

Distortion of DNA Origami on Graphene Imaged with Advanced TEM Techniques

While graphene may appear to be the ultimate support membrane for transmission electron microscopy (TEM) imaging of DNA nanostructures, very little is known if it poses an advantage over conventional carbon supports in terms of resolution and contrast. Microscopic investigations are carried out on DNA origami nanoplates that are supported...