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Robust 3D DNA origami can be used for the construction of artificial channels. Channels in the cell membrane transfer specific molecules and ions with a controlled gate system. DNA origami transmembrane channels were created and incorporated into the lipid bilayer [103]. This architecture consists of a robust cylinder‐like structure with a hollow interior and a short tubular pore that penetrates the lipid membrane (Figure 1.14a). To tightly attach to the lipid bilayer surface, multiple cholesterol molecules were incorporated at the bottom of the main body of the channel structure (Figure 1.14b). Using single‐channel electrophysiological measurements, the response of the origami channel showed properties similar to those of the natural ion channels in terms of conductance and channel gating. Using this channel, different lengths of single‐stranded DNAs with a stopper such as a stem and G‐quadruplex were discriminated by electrophysiological measurements. These results show that the origami channels can be further used to identify target molecules by detecting single‐molecule translocation.