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      Self-assembly of DNA into nanoscale three-dimensional shapes

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          Abstract

          Molecular self-assembly offers a ‘bottom-up’ route to fabrication with subnanometre precision of complex structures from simple components 1. DNA has proven a versatile building block 25 for programmable construction of such objects, including two-dimensional crystals 6, nanotubes 711, and three-dimensional wireframe nanopolyhedra 1217. Templated self-assembly of DNA 18 into custom two-dimensional shapes on the megadalton scale has been demonstrated previously with a multiple-kilobase ‘scaffold strand’ that is folded into a flat array of antiparallel helices by interactions with hundreds of oligonucleotide ‘staple strands’ 19, 20. Here we extend this method to building custom three-dimensional shapes formed as pleated layers of helices constrained to a honeycomb lattice. We demonstrate the design and assembly of nanostructures approximating six shapes — monolith, square nut, railed bridge, genie bottle, stacked cross, slotted cross — with precisely controlled dimensions ranging from 10 to 100 nm. We also show hierarchical assembly of structures such as homomultimeric linear tracks and of heterotrimeric wireframe icosahedra. Proper assembly requires week-long folding times and calibrated monovalent and divalent cation concentrations. We anticipate that our strategy for self-assembling custom three-dimensional shapes will provide a general route to the manufacture of sophisticated devices bearing features on the nanometer scale.

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          Most cited references24

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          Nucleic acid junctions and lattices.

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            Hierarchical self-assembly of DNA into symmetric supramolecular polyhedra.

            DNA is renowned for its double helix structure and the base pairing that enables the recognition and highly selective binding of complementary DNA strands. These features, and the ability to create DNA strands with any desired sequence of bases, have led to the use of DNA rationally to design various nanostructures and even execute molecular computations. Of the wide range of self-assembled DNA nanostructures reported, most are one- or two-dimensional. Examples of three-dimensional DNA structures include cubes, truncated octahedra, octohedra and tetrahedra, which are all comprised of many different DNA strands with unique sequences. When aiming for large structures, the need to synthesize large numbers (hundreds) of unique DNA strands poses a challenging design problem. Here, we demonstrate a simple solution to this problem: the design of basic DNA building units in such a way that many copies of identical units assemble into larger three-dimensional structures. We test this hierarchical self-assembly concept with DNA molecules that form three-point-star motifs, or tiles. By controlling the flexibility and concentration of the tiles, the one-pot assembly yields tetrahedra, dodecahedra or buckyballs that are tens of nanometres in size and comprised of four, twenty or sixty individual tiles, respectively. We expect that our assembly strategy can be adapted to allow the fabrication of a range of relatively complex three-dimensional structures.
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              Synthesis from DNA of a molecule with the connectivity of a cube.

              A principal goal of biotechnology is the assembly of novel biomaterials for analytical, industrial and therapeutic purposes. The advent of stable immobile nucleic acid branched junctions makes DNA a good candidate for building frameworks to which proteins or other functional molecules can be attached and thereby juxtaposed. The addition of single-stranded 'sticky' ends to branched DNA molecules converts them into macromolecular valence clusters that can be ligated together. The edges of these frameworks are double-helical DNA, and the vertices correspond to the branch points of junctions. Here, we report the construction from DNA of a covalently closed cube-like molecular complex containing twelve equal-length double-helical edges arranged about eight vertices. Each of the six 'faces' of the object is a single-stranded cyclic molecule, doubly catenated to four neighbouring strands, and each vertex is connected by an edge to three others. Each edge contains a unique restriction site for analytical purposes. This is the first construction of a closed polyhedral object from DNA.
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                Author and article information

                Journal
                0410462
                6011
                Nature
                Nature
                0028-0836
                1476-4687
                4 May 2009
                21 May 2009
                21 November 2009
                : 459
                : 7245
                : 414-418
                Affiliations
                [1 ] Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02115
                [2 ] Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115
                [3 ] Department of Genetics, Harvard Medical School, Boston, MA 02115
                [4 ] Department of Pathology, Harvard Medical School, Boston, MA 02115
                [5 ] Wyss Institute for Biologically Inspired Engineering, Harvard University, Cambridge, MA 02138
                Author notes
                Correspondence and requests for materials should be addressed to W.M.S. ( William_Shih@ 123456dfci.harvard.edu )
                Article
                nihpa105924
                10.1038/nature08016
                2688462
                19458720
                79238e99-e5eb-4675-b784-e308e6597f3f
                History
                Funding
                Funded by: Office of the Director : NIH
                Award ID: DP2 OD004641-01 ||OD
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