LAUSR

Efficient delivery of biomacromolecules or drugs across the cell membrane via endocytosis usually encounters inevitable entrapment in endosomes and subsequent degradation in lyso‐endosomes. To address this issue, a series of arginine‐rich cell penetrating polymers is designed and synthesized, which internalize into cells by inducing the formation of pores on the cell membrane, thereby crossing the cell membrane via direct translocation that fundamentally avoids endo/lysosomal entrapment. The structure−activity relationship studies show that PTn‐R2‐C6, which is a type of polymer that has two arginine residues and a flexible hexanoic acid linker in each side chain, exhibits excellent pore‐formation ability on the cell membrane. Further investigations indicate that PTn‐R2‐C6 rapidly transports plasmid DNAs into cytosol through a similar endocytosis‐independent pathway, thereby achieving significantly higher transfection efficiency and lower cytotoxicity than the gold‐standard transfection reagent PEI 25K. These results suggest the great potential of PTn‐R2‐C6 as a safe and efficient gene transfection reagent for wide applications including disease treatments, vaccine development, and biomedical research purposes.