A Smart DNA‐Based Nanosystem Containing Ribosome‐Regulating siRNA for Enhanced mRNA Transfection

Abstract Messenger RNA (mRNA) transfection is the prerequisite for the application of mRNA‐based therapeutics. In hard‐to‐transfect cells, such as macrophages, the effective transfection of mRNA remains a long‐standing challenge. Herein, a smart DNA‐based nanosystem is reported containing ribosome biogenesis‐promoting siRNA, realizing efficient mRNA transfection in macrophages. Four monomers are copolymerized to form a nanoframework (NF), including N ‐isopropylacrylamide (NIPAM) as the skeleton and acrydite‐DNA as the initiator to trigger the cascade assembly of DNA hairpins (H1‐polyT and H2‐siRNA). By virtue of the phase transition characteristic of polymeric NIPAM, below the lower critical solution temperature (LCST, ≈34 °C), the NF swells to expose polyT sequences to hybridize with the polyA tail of mRNA. Above the LCST, the NF deswells to encapsulate mRNA. The disulfide bond in the NF responds to glutathione, triggering the disassembly of the nanosystem; the siRNA and mRNA are released in response to triphosadenine and RNase H. The siRNA down‐regulates the expression of heat shock protein 27, which up‐regulates the expression of phosphorylated ribosomal protein S6. The nanosystem shows satisfactory mRNA transfection and translation efficiency in a mouse model. It is envisioned that the DNA‐based nanosystem will provide a promising carrier to deliver mRNA in hard‐to‐transfect cells and promote the development of mRNA‐based therapeutics.