Nanoparticles were formulated using a microfluidic device as previously described19. Luciferase mRNAs were diluted in
cholesterols and helper lipids (1,2-dioleoyl-sn-glycero-3-phosphoethanolamine, 1,2-distearoyl-sn-glycero-3-phosphocholine, 1,2-dioleoyl-3-trimethylammonium-propane, 1,2-di-O-octadecenyl-3-trimethylammonium propane dimethyldioctadecylammonium) were diluted in 100% ethanol.
PEGs, cholesterols, and helper lipids were purchased from Avanti Lipids. Citrate and ethanol phases were combined in a microfluidic device using syringes (Hamilton Company) at a flow rate of 3:1
Lokugamage, M.P., Vanover, D., Beyersdorf, J., Hatit, M.Z., Rotolo, L., Echeverri, E.S., Peck, H.E., Ni, H., Yoon, J.K., Kim, Y. and Santangelo, P.J., 2021. Optimization of lipid nanoparticles for the delivery of nebulized therapeutic mRNA to the lungs. Nature biomedical engineering, 5(9), pp.1059-1068.