Image-based analysis of lipid nanoparticle–mediated siRNA delivery, intracellular trafficking and endosomal escape
Abstract
Delivery of short interfering RNAs (siRNAs) remains a key challenge in the development of RNA interference (RNAi) therapeutics. A better understanding of the mechanisms of siRNA cellular uptake, intracellular transport and endosomal release could critically contribute to the improvement of delivery methods. Here we monitored the uptake of lipid nanoparticles (LNPs) loaded with traceable siRNAs in different cell types in vitro and in mouse liver by quantitative fluorescence imaging and electron microscopy. We found that LNPs enter cells by both constitutive and inducible pathways in a cell type-specific manner using clathrin-mediated endocytosis as well as macropinocytosis. By directly detecting colloidal-gold particles conjugated to siRNAs, we estimated that escape of siRNAs from endosomes into the cytosol occurs at low efficiency (1-2%) and only during a limited window of time when the LNPs reside in a specific compartment sharing early and late endosomal characteristics. Our results provide insights into LNP-mediated siRNA delivery that can guide development of the next generation of delivery systems for RNAi therapeutics.
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Following cellular internalization, lipid nanoparticles are usually trapped in endosomal compartments.
“Following cellular internalization, lipid nanoparticles are usually trapped in endosomal compartments137–139.”
From Lipid nanoparticles for mRNA delivery (2021) · cites this paper as [138]
Only a small amount of lipid nanoparticles may be able to escape from the endosome.
“Indeed, only a small amount of lipid nanoparticles may be able to escape from the endosome137–139.”
From Lipid nanoparticles for mRNA delivery (2021) · cites this paper as [138]
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- Authors
- Gilleron, J., Querbes, W., Zeigerer, A., Borodovsky, A., Marsico, G., Schubert, U., Manygoats, K., Seifert, S., Andree, C., Stöter, M., Epstein-Barash, H., Zhang, L., Koteliansky, V., Fitzgerald, K., Fava, E., Bickle, M., Kalaidzidis, Y., Akinc, A., Maier, M., & Zerial, M.
- Affiliations
- Alnylam Pharmaceuticals (United States) (US)
- Max Planck Institute of Molecular Cell Biology and Genetics (DE)
- German Center for Neurodegenerative Diseases (DE)
- Lomonosov Moscow State University (RU)
- Corresponding author
- Zerial, Marino
- ORCID
- Venue
- Nature Biotechnology
- Published
- 2013-06-23
- Volume
- 31
- Issue
- 7
- Pages
- 638-646
- ISSN
- 0733-222X (ISSN-L)
- 1087-0156
- 1546-1696
- OpenAlex
- W2155239981
- Cited by
- 1,681
- References
- 48
- Citations by year
- 2026: 248 · 2025: 229 · 2024: 175 · 2023: 136 · 2022: 125 · 2021: 140 · 2020: 106 · 2019: 98 · 2018: 89 · 2017: 87 · 2016: 93 · 2015: 81 · 2014: 55 · 2013: 17
- FWCI
- 36.70
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Biotechnology (publishedVersion)
- PubMed (publishedVersion)
- Max Planck Institute for Plasma Physics (submittedVersion)
- Site cant be reached (submittedVersion)
- DZNE Pub (submittedVersion)
- MPG.PuRe (Max Planck Society) (submittedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA Interference and Gene Delivery
- Physical Sciences › Materials Science › Biomaterials › Nanoparticle-Based Drug Delivery
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
- Keywords
- lipid nanoparticles, siRNA delivery, RNA interference therapy, intracellular trafficking, endosomal escape, clathrin-mediated endocytosis, macropinocytosis, colloidal gold particles, liver
- MeSH terms
- Animals, Endocytosis, Gold, HeLa Cells, Humans, Lipids, Microscopy, Electron, Gene Transfer Techniques, RNA, Small Interfering, Green Fluorescent Proteins, Mice, Metal Nanoparticles
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