Cells use membrane proteins as gatekeepers to transport ions and molecules, catalyze reactions, relay signals, and interact with other cells. DNA nanostructures with lipidic anchors are promising as membrane protein mimics because of their high tuneability. However, the design features specifying DNA nanostructure's functions in lipid membranes are yet to be fully understood. Here, we show that altering patterns of cholesterol units on a cubic DNA scaffold dramatically changes its interaction mode with lipid membranes. This results in simple design rules that allow a single DNA nanostructure to reproduce multiple membrane protein functions: peripheral anchoring, nanopore behavior and conformational switching to reveal membrane-binding units. Strikingly, the DNA-cholesterol cubes constitute the first open-walled DNA nanopores, as only a quarter of their wall is made of DNA. This functional diversity can increase our fundamental understanding of membrane phenomena, and results in sensing, drug delivery and cell manipulation tools. ASSOCIATED CONTENT Supporting Information. This material is available free of charge via the Internet at http://pubs.acs.org. DNA cage design and assembly, lipid vesicle preparation, membrane-binding study, dye-influx assay and molecular dynamic simulations.

This protocol provides a detailed guide for the design and assembly of membrane-spanning DNA nanopores and includes assays for characterizing channel function.TWEET A new protocol for design, assembly and characterisation of membrane-spanning DNA nanopores. COVER TEASER Design and characterisation of DNA nanoporesPlease indicate up to four primary research articles where the protocol has been used and/or developed.

A novel coumarin photocage with long-wavelength and high photolysis quantum yield shows solvent dependent photolysis.