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. Author manuscript; available in PMC: 2023 Jul 1.
Published in final edited form as: Nat Protoc. 2022 Oct 31;18(1):265–291. doi: 10.1038/s41596-022-00755-x

Fig. 1 |. Experimental design for preparation of liver, lung and spleen SORT LNPs.

Fig. 1 |

a, Inclusion of a SORT molecule in traditional four-component LNPs (which consist of ionizable cationic lipids, amphipathic phospholipids, cholesterol and PEG lipids) systematically alters the in vivo delivery profile of the resulting five-component SORT LNPs to enable tissue-specific delivery of mRNA to the liver, lungs and spleen of mice after IV administration. b, Chemical structures of the lipids used in this protocol, including the ionizable cationic lipids 4A3-SC8 and DLin-MC3-DMA (MC3), the phospholipids DOPE and DSPC, cholesterol, DMG-PEG and the SORT molecules DOTAP, 18PA and DODAP. c–e, Three mixing methods are described to prepare SORT LNP formulations introduced in this protocol: pipette mixing (c), vortex mixing (d) and microfluidic mixing methods (e). a adapted from ref.22, Springer Nature Limited. Images in c–e created with BioRender.com.