ATP-responsive polyethylenimine-based zinc-ligand nanoparticles for gene delivery

Abstract A zinc-coordinated ATP stimulus-responsive nanostructure was developed for CRISPR gene delivery. The system was based on low-molecular-weight polyethylenimine (PEI 1.8k ) attached to phenylboronic acid (PBA) and a pyridine zinc chelate. The nanostructure could form a polyplex with p-CRISPR DNA. The aim was for the PBA would bind to the ribose moiety of ATP and become hydrophilic, thus releasing the plasmid cargo. The in vitro findings demonstrated that the combination of a Zn-coordinated pyridine ligand, an ATP-sensitive PBA moiety, and a cationic backbone could act synergistically to condense DNA, enhance cellular internalization, disrupt endosomes effectively, and resist serum protein binding. This delivery platform enabled efficient CRISPR plasmid transfection in HEK 293T cells, even at low plasmid concentrations and under serum conditions ranging from 5% to 30%, outperforming the commonly used PEI 25k. We suggest that this novel method has the potential to be an appropriate non-viral vector for CRISPR/Cas9 gene editing in vivo and that it may potentially have therapeutic significance.