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.
journal_name
Nat Biotechnoljournal_title
Nature biotechnologyauthors
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 Mdoi
10.1038/nbt.2612subject
Has Abstractpub_date
2013-07-01 00:00:00pages
638-46issue
7eissn
1087-0156issn
1546-1696pii
nbt.2612journal_volume
31pub_type
杂志文章abstract：:A human skin allograft injury model in immunodeficient mice, engrafted with human peripheral blood mononuclear cells from a different donor, has been used to test whether reagents that block human T cell CD2 interactions with its principal ligand, LFA-3 (CD58), can inhibit immune reactions in vivo. In this model, huma...
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