From DNA origami to nanostructured mRNA-based delivery platforms.

DNA origami technique has established programmable nucleic acid nanostructures as a promising platform for biomedical applications. Incorporating RNA as the origami scaffold extends this potential into direct cytoplasmic activity, enabling transient protein expression without nuclear entry. However, RNA's distinct molecular structure and chemical lability impose design constraints with no equivalent in DNA-only systems, and the criteria for achieving robust intracellular gene expression from these hybrid constructs remain incompletely understood. This review synthesizes the current design principles governing mRNA-DNA hybrid origami assembly, including helicity adjustment, crossover geometry and folding condition optimization. It also examines the structural requirements for translational competence, where staple coverage of the 5' UTR eliminates expression and endosomal escape remains the primary delivery bottleneck. While hybrid origami systems have advanced, their immunological profile remains uncharacterized, and manufacturing scalability is unaddressed. Recent evidence that mRNA compaction alone can substantially enhance protein production in vivo suggests that scaffold-directed folding can offer advantages beyond nuclease protection. Establishing design rules that co-optimize structural integrity, translation competence, and immunological compatibility will be essential for translating mRNA scaffolded structures toward therapeutic applications.