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Imaging

Imaging to quantify lipid nanoparticle (LNP)-mediated delivery of small interfering RNAs

Combination of 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.

SciBX 6(30); doi:10.1038/scibx.2013.810
Published online Aug. 8, 2013

Unpatented; licensing status not applicable

Gilleron, J. et al. Nat. Biotechnol.; published online June 23, 2013;
doi:10.1038/nbt.2612
Contact: Marino Zerial, Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany
e-mail:
zerial@mpi-cbg.de