Precisely assembled DNA nanostructures are promising candidates for the delivery of biomolecule-based therapeutics. Herein, we introduce a facile strategy for the construction of a branched DNA-based nanoplatform for codelivery of gene editing (sgRNA/Cas9, targeting DNA in the nucleus) and gene silencing (antisense, targeting mRNA in the cytoplasm) components for synergistic tumor therapy in vitro and in vivo. In our design, the branched DNA structure can efficiently load a sgRNA/Cas9/antisense complex targeting a tumor-associated gene, PLK1, through DNA self-assembly. With the incorporation of an active targeting aptamer and an endosomal escape peptide by host–guest interaction, the biocompatible DNA nanoplatform demonstrates efficient inhibition of tumor growth without apparent systemic toxicity. This multifunctional DNA nanocarrier provides a new strategy for the development of gene therapeutics.

Chemically modified DNAh as been widely developed to fabricate various nucleic acid nanostructures for biomedical applications.Herein, we report afacile strategy for construction of branched antisense DNAand small interfering RNA(siRNA) co-assembled nanoplatform for combined gene silencing in vitro and in vivo.I no ur design, the branched antisense can efficiently capture siRNAw ith 3' overhangs through DNA-RNAhybridization. After being equipped with an active targeting group and an endosomal escape peptide by host-guest interaction, the tailored nucleic acid nanostructure functions efficiently as both delivery carrier and therapeutic cargo,w hich is released by endogenous RNase Hd igestion. The multifunctional nucleic acid nanosystem elicits an efficient inhibition of tumor growth based on the combined gene silencing of the tumor-associated gene polo-like kinase 1(PLK1). This biocompatible nucleic acid nanoplatform presents an ew strategy for the development of gene therapy.

DNA nanotechnology has been employed in the construction of self‐assembled nano‐biomaterials with uniform size and shape for various biological applications, such as bioimaging, diagnosis, or therapeutics. Herein, recent successful efforts to utilize multifunctional DNA origami nanoplatforms as drug‐delivery vehicles are reviewed. Diagnostic and therapeutic strategies based on gold nanorods, chemotherapeutic drugs, cytosine–phosphate–guanine, functional proteins, gene drugs, and their combinations for optoacoustic imaging, photothermal therapy, chemotherapy, immunological therapy, gene therapy, and coagulation‐based therapy are summarized. The challenges and opportunities for DNA‐based nanocarriers for biological applications are also discussed.