Advanced

Visualizing lipid-formulated siRNA release from endosomes and target gene knockdown.

Wittrup, Anders LU ; Ai, Angela; Liu, Xing; Hamar, Peter; Trifonova, Radiana; Charisse, Klaus; Manoharan, Muthiah; Kirchhausen, Tomas and Lieberman, Judy (2015) In Nature Biotechnology 33(8). p.870-870
Abstract
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... (More)
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. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Nature Biotechnology
volume
33
issue
8
pages
870 - 870
publisher
Nature Publishing Group
external identifiers
  • pmid:26192320
  • wos:000359274900027
  • scopus:84938898581
ISSN
1546-1696
DOI
10.1038/nbt.3298
language
English
LU publication?
yes
id
2a668074-f887-49ba-916c-f847fa3d2404 (old id 7726076)
alternative location
http://www.ncbi.nlm.nih.gov/pubmed/26192320?dopt=Abstract
date added to LUP
2015-08-10 12:51:29
date last changed
2017-11-19 03:20:07
@article{2a668074-f887-49ba-916c-f847fa3d2404,
  abstract     = {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 &lt;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.},
  author       = {Wittrup, Anders and Ai, Angela and Liu, Xing and Hamar, Peter and Trifonova, Radiana and Charisse, Klaus and Manoharan, Muthiah and Kirchhausen, Tomas and Lieberman, Judy},
  issn         = {1546-1696},
  language     = {eng},
  number       = {8},
  pages        = {870--870},
  publisher    = {Nature Publishing Group},
  series       = {Nature Biotechnology},
  title        = {Visualizing lipid-formulated siRNA release from endosomes and target gene knockdown.},
  url          = {http://dx.doi.org/10.1038/nbt.3298},
  volume       = {33},
  year         = {2015},
}