Visualizing lipid-formulated siRNA release from endosomes and target gene knockdown
By Anders Wittrup, Angela Ai, Xing Liu, P. Hamar, R. Trifonova, K. Charissé, M. Manoharan, T. Kirchhausen, J. Lieberman. Nature Biotechnology, 2015
602 citations recorded by the source corpus.
A central hurdle in developing small interfering RNAs (siRNAs) as therapeutics is the inefficiency of their delivery across the plasma and endosomal membranes to the cytosol, where they interact with the RNA interference machinery. With the aim of improving endosomal release, a poorly understood and inefficient process, we studied the uptake and cytosolic release of siRNAs, formulated in lipoplexes or lipid nanoparticles, by live-cell imaging and correlated it with knockdown of a target GFP reporter. siRNA release occurred invariably from maturing endosomes within ∼5–15 min of endocytosis. Cytosolic galectins immediately recognized the damaged endosome and targeted it for autophagy. However, inhibiting autophagy did not enhance cytosolic siRNA release. Gene knockdown occurred within a few hours of release and required <2,000 copies of cytosolic siRNAs. The ability to detect cytosolic release of siRNAs and understand how it is regulated will facilitate the development of rational strategies for improving the cytosolic delivery of candidate drugs.
Subfields: Genetics, Molecular Biology
View Paper: https://www.semanticscholar.org/paper/63fece0458c3e147917cc1ebe199b1e37bbfb8e2
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Citations in the corpus
- Concepts of nanoparticle cellular uptake, intracellular trafficking, and kinetics in nanomedicine. (2019, background): “For instance, in a study by Wittrup et al, cationic lipid nanoparticles formulated with siRNA were shown to only release their payload within ~10 minutes post-endocytosis in vitro in HeLa cells [249].”
- Brief update on endocytosis of nanomedicines (2019, background): “This process can sequester cytosolically localized nanocarriers and even engulf a leaky endosome, further impeding delivery [11].”
- The delivery of therapeutic oligonucleotides (2016, background): “A more recent study using highly sensitive fluorescence microscopy techniques documented release of siRNA from individual endosomes (182).”