Nucleic acid nanostructure platform for programming immune stimulation

A nucleic acid nanostructure comprising

• (i) a pentagonal bipyramid with 84 base pairs per edge, or an icosahedron with 42 base pairs per edge;
• and
• (ii) two or more, up to 40 copies of single stranded DNA including an immunostimulatory CpG motif bound to, or otherwise associated with the-nanostructure,
• wherein the distance between adjacent copies of the immunostimulatory CpG motif is from 1 to 100 Angstroms, inclusive, and
• wherein the number of copies of the immunostimulatory CpG motif, the distance between adjacent copies of the immunostimulatory CpG motifs, the location of the immunostimulatory CpG motifs on the nanostructure, the rigidity/flexibility of the single stranded DNA including a immunostimulatory CpG motif, the dimensionality of the immunostimulatory CpG motifs, the topology of the nanostructure, the ultra-structural organization of the nanostructure, the geometric shape of the nanostructure, or a combination thereof improves an immune response induced by the immunostimulatory CpG motifs relative to a control,
• wherein the control comprises the same nucleic acid nanostructure comprising a single copy of the immunostimulatory CpG,
• wherein the nanostructure comprises a single stranded nucleic acid scaffold sequence and one or more single stranded nucleic acid staple strands that hybridize to the scaffold sequence to form the three-dimensional nanostructure having a defined geometric shape, and
• wherein the two or more copies of single stranded DNA including an immunostimulatory CpG motif are located within one or more single stranded nucleic acid overhang extending from the 3′ or 5′ end of one or more selected staple strand of the nanostructure.