A study explores the differences in cationic cholesterol-dependent and cationic helper lipid-dependent tropism in lipid nanoparticles (LNPs) for mRNA delivery to the lung, liver, and heart. The research tested 196 LNPs delivering mRNA to 22 cell types across five tissues, revealing that cationic cholesterol led to a different lung:liver delivery ratio than cationic helper lipids. Combining cationic cholesterol with a cationic helper lipid enabled mRNA delivery to the heart and various lung cell types, including stem cell-like populations. The study highlights the utility of charge-dependent LNP tropism for targeting multiple tissues. LNPs with cationic cholesterol showed increased liver delivery compared to those with cationic helper lipids, suggesting that charge-dependent tropism may vary based on the charged component. The study also identified that LNP $ ^{++} $, containing both cationic cholesterol and cationic helper lipid, delivered mRNA to heart endothelial cells and various lung cell types, including nonendothelial cells. These findings suggest that charge-dependent tropism can be used to target multiple tissues, including the heart, liver, and lung, for genetic diseases. The study also emphasizes the importance of understanding the biological mechanisms behind these differences and the need for further research to validate findings in non-human primates. The results demonstrate the potential of charge-dependent LNP tropism for improving mRNA delivery to multiple tissues, which could be useful for treating genetic disorders affecting multiple organs.A study explores the differences in cationic cholesterol-dependent and cationic helper lipid-dependent tropism in lipid nanoparticles (LNPs) for mRNA delivery to the lung, liver, and heart. The research tested 196 LNPs delivering mRNA to 22 cell types across five tissues, revealing that cationic cholesterol led to a different lung:liver delivery ratio than cationic helper lipids. Combining cationic cholesterol with a cationic helper lipid enabled mRNA delivery to the heart and various lung cell types, including stem cell-like populations. The study highlights the utility of charge-dependent LNP tropism for targeting multiple tissues. LNPs with cationic cholesterol showed increased liver delivery compared to those with cationic helper lipids, suggesting that charge-dependent tropism may vary based on the charged component. The study also identified that LNP $ ^{++} $, containing both cationic cholesterol and cationic helper lipid, delivered mRNA to heart endothelial cells and various lung cell types, including nonendothelial cells. These findings suggest that charge-dependent tropism can be used to target multiple tissues, including the heart, liver, and lung, for genetic diseases. The study also emphasizes the importance of understanding the biological mechanisms behind these differences and the need for further research to validate findings in non-human primates. The results demonstrate the potential of charge-dependent LNP tropism for improving mRNA delivery to multiple tissues, which could be useful for treating genetic disorders affecting multiple organs.