Towards a bottom-up reconstitution of the nuclear pore complex

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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, as such, one can not directly see or experience any of the events that occur down there. The long answer can be found in this thesis. Here, I present and discuss a wide array of nanopores, from biological nanopores like the nuclear pore complex (NPC), to solidstate nanopores, and DNA-origami nanopores. While the central focus of my research is to understand the inner workings of the NPC, a short journey into the world of ion transport in solid-state nanopores is first undertaken, with special emphasis on the random fluctuations of the ion flow within the nanopore, referred to as current noise. Next, I introduce the concept of biomimetic nanopores, where a solid-state nanopore is ‘camouflaged’ by coating its inner surface with purified proteins, resulting in an entity that behaves somewhat like a real NPC. Biomimetic nanopores have enabled us to mimic, study, and gain new insights into how the real NPC works, and bear great potential for further developments and discoveries.