Fluorophore-Tagged Poly-Lysine RAFT Agents: Controlled Synthesis of Trackable Cell-Penetrating Peptide–Polymers

[Image: see text] The conjugation of a fluorophore and a variety of cell-penetrating peptides onto a RAFT agent allowed for the synthesis of polymers of defined sizes with quantifiable cell-uptake. Each peptide–RAFT agent was used to polymerize acrylamide, acrylate, and styrene monomers to form high or low molecular weight polymers (here 50 or 7.5 kDa) with the peptide having no influence on the RAFT agent’s control. The incorporation of a single fluorophore per polymer chain allowed cellular analysis of the uptake of the size-specific peptide–polymers via flow cytometry and confocal microscopy. The cell-penetrating peptides had a direct effect on the efficiency of polymer uptake for both high and low molecular weight polymers, demonstrating the versatility of the strategy. These “all-in-one”, synthetically accessible RAFT agents allow highly controlled preparation of synthetic peptide–polymer conjugates and subsequent quantification of their delivery into cells.