1.D.203.  The DNA-Origami-Protein Nanopore (DOP-NP) Family 

Functionalized DNA-origami-protein nanopores generate large transmembrane channels with programmable size-selectivity (Shen et al. 2023). The DNA-origami technique has enabled the engineering of transmembrane nanopores with programmable size and functionality, showing promise for building biosensors and synthetic cells. Shen et al. 2023 took advantage of pneumolysin (PLY; TC# 1.C.12.1.5), a bacterial toxin that potently forms wide ring-like channels in cell membranes, to construct hybrid DNA-protein nanopores. This PLY-DNA-origami complex, in which a DNA-origami ring corrals up to 48 copies of PLY, targets the cholesterol-rich membranes of liposomes and red blood cells, readily forming uniformly sized pores with an average inner diameter of approximately 22 nm. Such hybrid nanopores facilitate the exchange of macromolecules between perforated liposomes and their environment, with the exchange rate negatively correlating with the macromolecule size (diameters of gyration: 8-22 nm). The DNA ring can be decorated with intrinsically disordered nucleoporins to further restrict the diffusion of traversing molecules, highlighting the programmability of the hybrid nanopores. PLY-DNA pores provide an enabling biophysical tool for studying the cross-membrane translocation of ultralarge molecules and open new opportunities for analytical chemistry, synthetic biology, and nanomedicine (Shen et al. 2023).