DNA origami encoding for gene expression and contransfection

A nucleic acid nanostructure for expression of a target gene in a eukaryotic cell comprising:

• (a) a single-stranded DNA (ssDNA) scaffold strand that comprises an expression cassette comprising at least one nucleic acid sequence encoding the target gene, a promoter, a terminator, and a polyadenylation signal sequence, and
• (b) at least one enhancer staple that comprises a sequence that binds to the ssDNA scaffold strand and upon binding enhances expression of the target gene when the nucleic acid nanostructure is inserted into the eukaryotic cell, as compared to a nucleic acid nanostructure without the enhancer staple,
• wherein the at least one enhancer staple binds to the ssDNA scaffold strand at a start region of the promoter and at the polyadenylation signal sequence;
• whereby when the at least one enhancer staple binds to both the start region of the promoter and the polyadenylation signal sequence, the single enhancer staple acts as a splint that brings together the start region of the promoter and the polyadenylation signal sequence.