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@ARTICLE{Gilleron:136981,
author = {Gilleron, Jerome and Querbes, William and Zeigerer, Anja
and Borodovsky, Anna and Marsico, Giovanni and Schubert,
Undine and Manygoats, Kevin and Seifert, Sarah and Andree,
Cordula and Stöter, Martin and Epstein-Barash, Hila and
Zhang, Ligang and Koteliansky, Victor and Fitzgerald, Kevin
and Fava, Eugenio and Bickle, Marc and Kalaidzidis, Yannis
and Akinc, Akin and Maier, Martin and Zerial, Marino},
title = {{I}mage-based analysis of lipid nanoparticle-mediated
si{RNA} delivery, intracellular trafficking and endosomal
escape.},
journal = {Nature biotechnology},
volume = {31},
number = {7},
issn = {1087-0156},
address = {New York, NY},
publisher = {Nature America},
reportid = {DZNE-2020-03303},
pages = {638-646},
year = {2013},
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.},
keywords = {Animals / Endocytosis: genetics / Gene Transfer Techniques
/ Gold: administration $\&$ dosage / Gold: chemistry / Green
Fluorescent Proteins: antagonists $\&$ inhibitors / Green
Fluorescent Proteins: genetics / HeLa Cells / Humans /
Lipids: administration $\&$ dosage / Lipids: chemistry /
Lipids: genetics / Metal Nanoparticles: administration $\&$
dosage / Metal Nanoparticles: chemistry / Mice / Microscopy,
Electron / RNA, Small Interfering: administration $\&$
dosage / RNA, Small Interfering: chemistry / RNA, Small
Interfering: genetics / Lipids (NLM Chemicals) / RNA, Small
Interfering (NLM Chemicals) / Green Fluorescent Proteins
(NLM Chemicals) / Gold (NLM Chemicals)},
cin = {AG Fava 1},
ddc = {660},
cid = {I:(DE-2719)1013016},
pnm = {342 - Disease Mechanisms and Model Systems (POF3-342)},
pid = {G:(DE-HGF)POF3-342},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:23792630},
doi = {10.1038/nbt.2612},
url = {https://pub.dzne.de/record/136981},
}