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bioRxiv · 10.1101/2020.12.02.407601

Illuminating endosomal escape of polymorphic lipid nanoparticles that boost mRNA delivery

Herrera, M.·Kim, J.·Eygeris, Y.·Jozic, A.·Sahay, G.
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Abstract

Lipid-based nanoparticles (LNPs) for the delivery of mRNA have jumped to the forefront of non-viral gene delivery. Despite this exciting development, poor endosomal escape after LNP cell entry remains an unsolved, rate-limiting bottleneck. Here we report the use of galectin 8-GFP (Gal8-GFP) cell reporter system to visualize the endosomal escaping capabilities of LNPs encapsulating mRNA. LNPs substituted with phytosterols in place of cholesterol exhibited various levels of Gal8 recruitment in the Gal8-GFP reporter system. In live-cell imaging, the results of the LNPs containing {beta}-sitosterol (LNP-Sito) showed a 10-fold increase in detectable endosomal perturbation events when compared to the standard cholesterol LNPs (LNP-Chol), suggesting the superior capability of LNP-Sito to escape from endosomal entrapment. Trafficking studies of these LNPs showed strong localization with late endosomes. This highly sensitive and robust Gal8-GFP reporter system can be a valuable tool to elucidate intricacies of LNP trafficking and ephemeral endosomal escape events, enabling the advancement in gene delivery.

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Herrera, M., Kim, J., Eygeris, Y., Jozic, A., Sahay, G.. 2020-12-03. Illuminating endosomal escape of polymorphic lipid nanoparticles that boost mRNA delivery. https://doi.org/10.1101/2020.12.02.407601

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