Protein and mRNA Delivery Enabled by Cholesteryl‐Based Biodegradable Lipidoid Nanoparticles

Developing safe and efficient delivery systems for therapeutic biomacromolecules is a long‐standing challenge. Herein, we report a newly developed combinatorial library of cholesteryl‐based disulfide bond‐containing biodegradable cationic lipidoid nanoparticles. We have identified a subset of this library which is effective for protein and mRNA delivery in vitro and in vivo. These lipidoids showed comparable transfection efficacies but much lower cytotoxicities compared to the Lpf2k in vitro. In vivo studies in adult mice demonstrated the successful delivery of genome engineering protein and mRNA molecules in the skeletal muscle (via intramuscular injection), lung and spleen (via intravenous injection), and brain (via lateral ventricle infusion). Intracellular delivery: a new combinatorial library of cholesteryl‐based and reduction‐responsive lipidoid nanoparticles can deliver genome engineering protein and mRNA molecules in vitro and in vivo. Using adult Ai14 mouse model, successful Cre‐mediated gene recombination events were observed in skeletal muscle, brain, lung, and spleen through the local and systemic administrations.