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Encapsulation state of messenger RNA inside lipid nanoparticles
Authors:Mark L. Brader  Sean J. Williams  Jessica M. Banks  Wong H. Hui  Z. Hong Zhou  Lin Jin
Affiliation:1. Moderna, Inc., Cambridge, Massachusetts;2. Department of Microbiology, Immunology, and Molecular Genetics, Los Angeles, Los Angeles, California;3. California NanoSystems Institute, University of California, Los Angeles, Los Angeles, California
Abstract:
Understanding the structure of messenger RNA (mRNA) lipid nanoparticles, and specifically the microenvironment of the mRNA molecules within these entities, is fundamental to advancing their biomedical potential. Here, we show that a permeating cationic dye, thionine, can serve as a cryogenic electron microscopy contrasting agent by binding selectively to encapsulated mRNA without disturbing lipid nanoparticle morphology. Cryo-electron microscopy images identify the mRNA location, revealing that mRNA may exist within solvent-filled cavities or may be substantially lipid associated.
Keywords:
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