LNPs were formulated via microfluidic mixing using a Precision Nanosystems NanoAssemblr Ignite instrument with a formulation were composed of Dlin-MC3-DMA/DSPC/DMG-PEG2k/Cholesterol at a molar ratio of (50/10/1.5/38.5) with an N:P ratio of 6. Lipids were solubilized in ethanol. The RNA cargo was comprised of a 1:1 molar ratio of mRNA reporter cargo and formulated in a 50mM citrate buffer at pH 3.0. After mixing, nanoparticles were transferred to G2 dialysis cassettes (10k MWCO) and dialyzed into PBS in at least 200x excess buffer for two hours at room temperature followed by overnight dialysis with gentle stirring in refreshed buffer at 4°C. LNP size and polydispersity were determined using a Zetasizer Ultra Malvern Panalytical instrument and are presented in Figure .