Carbon nanotubes (CNTs) can be used to create unique fluidic systems for studying ion transport in nanochannels due to their well-defined geometry, atomically smooth and chemically inert surface, and similarity to transmembrane protein pores. Here, we report the reversible molecular gating of ion transport across DNA-functionalized CNT membranes. The diffusive transport rates of ferricyanide ions through membranes, each with an array of aligned transmembrane CNT channels, were monitored.Single-stranded DNA (T15) gate molecules were covalently linked to CNT channel entrances, and reversible opening and closing of CNT channels were achieved by the addition and removal of complementary DNAs (A15) with a remarkable open/close flux ratio of >1000, which is substantially higher than the protein-gated CNT systems reported previously. Furthermore, at least two-orders of magnitude difference in ion fluxes was observed when single base-pair mismatched DNAs were used in place of the complementary DNAs. Comprehensive theoretical analysis is also presented. The experimental results can be explained by steric hindrance, ion partitioning, and electrostatic repulsion at the CNT entrances, as well as the thermodynamics of DNA binding.Protein channels are fascinating structures, which can selectively and efficiently transport essential chemicals through cell membranes. The fabrication of synthetic membranes with nanopores that mimic biological transmembrane protein channels have numerous applications, ranging from drug delivery, water purication, and molecular sieving, to DNA sensing.1-4 These membranes possess selective gate chemistry at the pore entrance, a mechanism for fast hydrodynamic ow, and a mechanism to stimulate the channel.5 Several different approaches were developed to create biomimetic membranes. Previous studies have investigated the use of porous alumina or track-etched polycarbonate substrates with well-ordered nanoporous structures and selective chemistry.3,4,6-8 However, these works did not show the capability to create an efficient chemical layer to act as a gatekeeper over the pores and no enhancement to hydrodynamic ow was observed either. Recently, carbon nanotubes (CNTs) have been investigated as biomimetic uidic channels due to their fast hydrodynamic velocity proles, highly uniform and tunable pore diameters, and potential gating capabilities.9-14 Early molecular dynamics (MD) simulations predicted strong hydrogen bonding of water in hydrophobic CNTs, resulting in a faster hydrodynamic ow rate than that expected for conventional porous platforms 9 and on the same order as water through Aquaporin-1. MD simulations and experiments have shown that the increased hydrodynamic ow velocities through CNTs can be attributed to the atomic smoothness of the graphitic surface displaying near-perfect slip properties. 9,11 In other studies, a fast ow rate of molecules was also predicted based on the near frictionless nature of the CNT walls. 11,15,16