Fig. 5: LNP composition adjustment enabled simultaneous organ-targeted mRNA accumulation and translation.

a, b Typical four-component 6Ac1-C12 LNPs primarily accumulated in the liver post i.v. administration. Accumulation images (a) and quantification (b) of 6Ac1-C12 LNPs containing Cy5-mRNA were evaluated. c FACS was used to determine the mRNA distribution in hepatic cell types after treating 5Ac1-C12 and 6Ac1-C12 Liver LNPs. d The schematic showed the deletion of stop cassette by delivering Cre mRNA to Ai9 mice for activation of tdTomato expression. e Hepatic cell type transfection analysis of 5Ac1-C12 and 6Ac1-C12 Liver LNPs. Cholesterol removal from Liver LNPs decreased the liver specificity (Fluc mRNA, 0.25 mg kg⁻¹). Schematic (f), in vivo images (g), and targeting alteration (h) of 6Ac1-C12 Liver LNPs after removing the cholesterol component were shown. 3-Comp Lung LNPs without cholesterol and phospholipid demonstrated superior pulmonary targeting compared to 4- and 5-Comp Lung LNPs. The accumulation (i) and lung-targeting (j) of 6Ac1-C12 Lung LNPs with different components were assayed. k Cholesterol effect on lipoprotein adsorption of Lung LNPs by protein corona analysis. l Pulmonary cell type distribution analysis of Lung LNPs. m Quantification of pulmonary cell transfection by Lung LNPs. n Stability of 6Ac1-C12 3-Comp Lung LNPs. These LNPs remained stable post 30-d incubation. o 6Ac1-C12 3-Comp Lung LNPs were well tolerated in vivo. Alanine aminotransferase (ALT), aspartate aminotransferase (AST), blood urea nitrogen (BUN), and creatinine (CREA) were evaluated. All data are presented as mean±s.d. (n = 3 biologically independent animals). Statistical significance in h, k, o was calculated with a two-tailed unpaired t-test, and those in b, j were calculated using one-way ANOVA with Dunnett’s multiple comparisons test: ^****P < 0.0001; ^***P < 0.001; ^**P < 0.01; ^*P < 0.05; ns, no significant difference.