A Thermal and Enzymatic Dual‐Stimuli Responsive DNA‐Based Nanomachine for Controlled mRNA Delivery

Abstract

The extreme instability of mRNA makes the practical application of mRNA-based vaccines heavily rely on efficient delivery system and cold chain transportation. Herein, a DNA-based nanomachine, which achieves programmed capture, long-term storage without cryopreservation, and efficient delivery of mRNA in cells, is developed. The polythymidine acid (Poly-T) functionalized poly(N-isopropylacrylamide) (DNA-PNIPAM) is synthesized and assembled as the central compartment of the nanomachine. The DNA-PNIPAM nano-assembly exhibits reversible thermal-responsive dynamic property: when lower than the low critical solution temperature (LCST, ≈32 °C) of PNIPAM, the DNA-PNIPAM transforms into extension state to expose the poly-T, facilitating the hybridization with polyadenylic acid (Poly-A) tail of mRNA; when higher than LCST, DNA-PNIPAM re-assembles and achieves an efficient encapsulation of mRNA. It is remarkable that the DNA-PNIPAM nano-assembly realizes long-term storage of mRNA (≈7 days) at 37 °C. Biodegradable 2-hydroxypropyltrimethyl ammonium chloride chitosan is assembled on the outside of DNA-PNIPAM to facilitate the endocytosis of mRNA, RNase-H mediating mRNA release occurs in cytoplasm, and efficient mRNA translation is achieved. This work provides a new disign principle of nanosystem for mRNA delivery.

Cellular Trafficking of Nanotechnology‐Mediated mRNA Delivery

“…[11,[80][81][82][83] Of note, delivery vectors based on electrostatic loading approaches are afflicted by positive charge-related cytotoxicity and interference from negatively charged biomolecules. [11] To avoid these cationrelated side effects, non-electrostatic loading strategies driven by other interactions like base complementary pairing, [84][85][86] hydrogen bonding, [87] and coordination have been explored for mRNA delivery. [88,89] For example, forming metal/mRNA complex via metal coordination-mediated self-assembly is a practical strategy to condense mRNA.…”

“…With the development of nucleic acid self‐assembly technology, nucleic acid‐based nanostructures have been employed for mRNA delivery due to their good biocompatibility, facile modification, and structural programmability. [ 84,137 ,. 138 ] For instance, Yoshinaga et al.…”

Bioinspired Spatiotemporal Management toward RNA Therapies

“…The red blood cell extracellular vesicles demonstrated microRNA knockdown and gene knockout in leukemia and breast cancer cells both in vitro and in vivo , without cytotoxicity . Another study designed a spatiotemporal mRNA encapsulated DNA-PNIPAM based nanomachine achieved ∼7 days mRNA storage at 37 °C and efficient mRNA translation in DC 2.4 cells . Specifically, this nanomachine was designed with thermal and enzymatic dual-stimuli responsive properties, where mRNA with a Poly-A tail could hybridized with Poly-T on the nanomachine in a time-efficient manner when the temperature was below a low critical solution temperature (LCST, ≈32 °C).…”

Construction of Polymeric DNA Network and Application for Cell Manipulation

“…developed a thermal‐ responsive nanomachine by combining DNA with poly‐N‐isopropylacrylamide (PNIPAM), achieving an efficient mRNA encapsulation. [ 38 ] By taking full advantage of the programmability of DNA molecules and introducing silver nanoclusters, Geng et al . presented a novel fluorescent antibacterial DNA network based on RCA strategy.…”