Thursday, August 8, 2013
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Imaging to quantify lipid
nanoparticle (LNP)-mediated delivery of small interfering RNAs
high-resolution light and electron microscopy (EM) could be used to quantify
uptake and trafficking of LNP-delivered siRNAs. In cultured cells,
quantitative image analysis of cells treated with fluorescently labeled
GFP-targeting siRNAs revealed time-dependent siRNA uptake that corresponded
with silencing of GFP. In cells, EM and mathematical modeling revealed an
escape rate of 1%-2% for gold-labeled siRNAs from endosomes, corresponding to
2,000-4,000 molecules per cell. In cultured cells and in mice, small molecule-mediated
inhibition of endosomal maturation did not affect siRNA escape, indicating
that siRNAs escape from early and not mature endosomes. Next steps include
identifying small molecules that impact cellular uptake of siRNAs.
Published online Aug. 8, 2013
status not applicable
Gilleron, J. et al. Nat.
Biotechnol.; published online June 23, 2013;
Contact: Marino Zerial, Max Planck Institute of Molecular
Cell Biology and Genetics, Dresden, Germany
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