The formation of highly ordered chiral organic/ inorganic films with high density and long-range orientation is important in constructing chiral devices,s uch as broadband polarization devices,liquid-crystal displays,ornegative-reflection materials.Afeasible strategy is presented to fabricate three-dimensional mesostructured chiral DNA-silica assemblies into large-scale oriented arrangements.T he highly ordered film was aligned by am ica crystal substrate with the bridging effect of suitable divalent metal ions,followed by the growth of the DNA-silica composite by bottom-up assembly with a"quartet templating" method. This simple and effective route would perform well in the alignment and arrangement of highly charged biomolecules,s uch as polypeptides,p roteins, viruses,and their inorganic assemblies,and furthermore could allowthe fabrication of chiral optical materials with long-range ordering.The precise engineering of nanostructures over am acroscopic surface into highly ordered three-dimensional (3D) arrays is as ubstantial challenge in many applications in nanotechnology.[1] Among them, chiral nanostructures are of great interest.[2] Thes ignificant aspects of chiral devices, especially chiral films such as broadband polarizers, [3] chiral liquid crystals, [4] and chiral filters, [5] often arise from their long-range arrangement. Thetemplate-assisted self-assembly (TASA) method to guide suitable chiral molecules to assemble into 3D large arrays is an effective strategy to fabricate long-range ordered chiral films.The combination of top-down substrate design and bottom-up molecular selfassembly would enable us to integrate individual nanostructures into arranged functional devices. [6] As one of the most attractive biomolecular building blocks,D NA molecules provide high stereoregularity for building artificial nanostructures and possess inherent chirality to assemble into delicate chiral materials. [1b, 7] Using elaborate intermolecular interactions,D NA can position materials with precision into periodic or aperiodic structures.[8] Thus,controlling the orientation and arrangement of DNAassembles on supporting substrates through TASA may be the fine approach for the fabrication of highly ordered films with chiral structures.Previously,w es uccessfully oriented and selectively arranged achiral 2D mesostructured DNA-silica platelets on the surface of al ithographic silicon substrate through TASA.[9] Furthermore,chiral DNA-silica impellers have also been fabricated on silica wafers,s howing both absorptionand scattering-based optical activity (OA).[10] Unfortunately, the randomly distributed impellers suffer from weak OA, raising the requirements in terms of controlling the orientation of chiral DNA-silica assemblies.M eanwhile,t he lithographic process is energy-intensive,a nd the resolution is limited to 7nmlength scales.[11] It is necessary to choose anew substrate to align the structure.Despite commonly used substrates having organized lithographic patterns,f lat crystalline substrates (highly ...