Redox-Responsive Polymeric Nanogels as Efficient mRNA Delivery Vehicles in Caenorhabditis elegans

Efficient delivery of sensitive nucleic acid payloads, including mRNA, in Caenorhabditis elegans remains challenging, especially with traditional, labor-intensive transgenesis methods. We addressed these challenges using polymeric nanogels (NGs) as an advanced platform for mRNA delivery in C. elegans . These polymeric delivery vehicles can be engineered to suit desired applications owing to their chemical versatility, resulting from the ability to conjugate multiple functional groups onto the same backbone. Here, we validate the in vivo RNA delivery potential of redox-responsive NGs. The NGs showed up to 72.4 % RNA encapsulation and 6.61 % loading efficiencies and facilitated the controlled release of the mRNA payloads at intracellular concentrations of the reducing agent glutathione, where most of the RNA was released within 24 hours. As a proof of concept, we successfully delivered green fluorescent protein (GFP)-expressing mRNA using NGs in C. elegans for the first time. Physicochemical characterization revealed uniform NG size and charge, and fluorescence microscopy confirmed GFP expression in the gut after 24 hours of treatment. Our findings show NGs' potential as an mRNA delivery system in C. elegans .