The study investigates the impact of cationic cholesterol on the tropism of lipid nanoparticles (LNPs) for delivering mRNA to various tissues, including the lung, liver, and heart. The authors tested 196 LNPs with different combinations of charged cholesterols and helper lipids in mice, finding that charged cholesterols led to a different lung/liver delivery ratio compared to charged helper lipids. Specifically, LNPs containing cationic cholesterols and neutral helper lipids showed increased liver delivery, while those with cationic cholesterols and cationic helper lipids improved non-liver/liver tropism. Additionally, the combination of cationic cholesterol and cationic helper lipid resulted in mRNA delivery to the heart and several lung cell types, including stem cell-like populations. These findings highlight the complexity of LNP tropism and suggest that charged cholesterols can influence delivery differently from charged helper lipids. The study also emphasizes the utility of exploring charge-dependent LNP tropism for targeted mRNA delivery to specific tissues.The study investigates the impact of cationic cholesterol on the tropism of lipid nanoparticles (LNPs) for delivering mRNA to various tissues, including the lung, liver, and heart. The authors tested 196 LNPs with different combinations of charged cholesterols and helper lipids in mice, finding that charged cholesterols led to a different lung/liver delivery ratio compared to charged helper lipids. Specifically, LNPs containing cationic cholesterols and neutral helper lipids showed increased liver delivery, while those with cationic cholesterols and cationic helper lipids improved non-liver/liver tropism. Additionally, the combination of cationic cholesterol and cationic helper lipid resulted in mRNA delivery to the heart and several lung cell types, including stem cell-like populations. These findings highlight the complexity of LNP tropism and suggest that charged cholesterols can influence delivery differently from charged helper lipids. The study also emphasizes the utility of exploring charge-dependent LNP tropism for targeted mRNA delivery to specific tissues.