Linear ABC Amphiphilic Triblock Copolymers for Complexation and Protection of dsRNA

We herein report the synthesis and characterisation of linear ABC triblock copolymers, investigation of their self-assembly in aqueous solution, and complexation and protection with double stranded-RNA (dsRNA). The amphiphilic triblock copolymers were synthesised via reversible addition–fragmentation chain transfer (RAFT) polymerisation. The precisely controlled polymerisation allowed for modification of the degree of polymerisation of quaternised 2-(dimethylamino)ethyl methacrylate (QDMAEMA, Q), tert-butyl acrylamide (tBAA, B) and N,N-dimethyl acrylamide (DMA, D) blocks, tailoring hydrophobicity. The Q homopolymer was synthesised as a macromolecular chain-transfer agent. The cationic functionality provides the ability for electrostatic interaction of the triblock copolymers with anionic biomolecules, such as dsRNA, for therapeutic or agrochemical delivery applications. The B second block was designed to provide strong anchoring of the assembled structures for enhanced stability. As illustrated by 1H NMR spectroscopy, Q-b-B-b-D linear ABC triblock copolymers were prepared with molecular weights 30, 37 and 44 kDa. The self-assembly of these amphiphilic triblock copolymers in aqueous solution was confirmed by dynamic light scattering (DLS) and transmission electron microscopy (TEM). Furthermore, the potential of these tailored block copolymers as vehicles for dsRNA delivery was demonstrated through complexation and protection of the anionic biomolecule dsRNA against destabilisation at high salt concentration and enzymatic degradation by RNase A, confirmed by ethidium bromide exclusion and agarose gel electrophoresis assays.