Modification of DNA Nanostructures with Functional Peptides Through Copper-Free Click Chemistry

The provided protocol outlines a method for synthesizing a peptide-modified three-arm DNA nanostructure. The protocol details the functionalization of three partially complementary amino-modified single-stranded DNA molecules with azide-modified peptide, denoted "PeB," using copper-free click chemistry. This PeB peptide, derived from the neutralizing antibody HC19, effectively targets the influenza A virus. Subsequently, the peptide-functionalized DNA structures are assembled into a three-arm DNA structure, enabling multivalent presentation of the peptides to enhance binding and inhibit viral particles. While this protocol specifically describes the synthesis of simple trivalent DNA-peptide structures, the general approach can be scaled to decorate more complex DNA nanostructures such as those formed from tile- or origami-based architectures.