De novo design of semisynthetic protein nanopores

Abstract Protein nanopores are essential components of single-molecule oligonucleotide sequencing and sensing devices. Here, we demonstrate that installing additional de novo subunits enables large-scale architectural changes of nanopore complexes. We design de novo proteins that integrate seamlessly with the CsgG pore to form 18-subunit, 315-kilodalton complexes with precisely sculpted pore architectures and tailored ion conduction, opening new possibilities for engineering enhanced nanopores with customized structural and functional properties. Competing Interest Statement The authors declare the following competing financial interest: L.S, A.J.S, R.G.H, R.C.G, N.F.P, E.J.W, and W.F.D are co-inventors of the international patent application WO 2024/ 033447A1 entitled: De novo pores. A.J.S, R.G.H, R.C.G and E.J.W are members of Oxford Nanopore Technologies plc, which commercializes nanopore devices. W.F.D received financial support from Oxford Nanopore Technologies plc for this work.