TY  - JOUR
AU  - Gilleron, Jerome
AU  - Querbes, William
AU  - Zeigerer, Anja
AU  - Borodovsky, Anna
AU  - Marsico, Giovanni
AU  - Schubert, Undine
AU  - Manygoats, Kevin
AU  - Seifert, Sarah
AU  - Andree, Cordula
AU  - Stöter, Martin
AU  - Epstein-Barash, Hila
AU  - Zhang, Ligang
AU  - Koteliansky, Victor
AU  - Fitzgerald, Kevin
AU  - Fava, Eugenio
AU  - Bickle, Marc
AU  - Kalaidzidis, Yannis
AU  - Akinc, Akin
AU  - Maier, Martin
AU  - Zerial, Marino
TI  - Image-based analysis of lipid nanoparticle-mediated siRNA delivery, intracellular trafficking and endosomal escape.
JO  - Nature biotechnology
VL  - 31
IS  - 7
SN  - 1087-0156
CY  - New York, NY
PB  - Nature America
M1  - DZNE-2020-03303
SP  - 638-646
PY  - 2013
AB  - 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
KW  - Animals
KW  - Endocytosis: genetics
KW  - Gene Transfer Techniques
KW  - Gold: administration & dosage
KW  - Gold: chemistry
KW  - Green Fluorescent Proteins: antagonists & inhibitors
KW  - Green Fluorescent Proteins: genetics
KW  - HeLa Cells
KW  - Humans
KW  - Lipids: administration & dosage
KW  - Lipids: chemistry
KW  - Lipids: genetics
KW  - Metal Nanoparticles: administration & dosage
KW  - Metal Nanoparticles: chemistry
KW  - Mice
KW  - Microscopy, Electron
KW  - RNA, Small Interfering: administration & dosage
KW  - RNA, Small Interfering: chemistry
KW  - RNA, Small Interfering: genetics
KW  - Lipids (NLM Chemicals)
KW  - RNA, Small Interfering (NLM Chemicals)
KW  - Green Fluorescent Proteins (NLM Chemicals)
KW  - Gold (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:23792630
DO  - DOI:10.1038/nbt.2612
UR  - https://pub.dzne.de/record/136981
ER  -