Journal
BIOMATERIALS SCIENCE
Volume 9, Issue 12, Pages 4289-4300Publisher
ROYAL SOC CHEMISTRY
DOI: 10.1039/d0bm01947j
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Funding
- NEI NIH HHS [R21 EY031066] Funding Source: Medline
- NHLBI NIH HHS [R01 HL146736] Funding Source: Medline
- NIGMS NIH HHS [U24 GM129547] Funding Source: Medline
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The study utilized a Galectin 8-GFP cell reporter system to visualize the endosomal escape capabilities of lipid-based nanoparticles (LNPs) encapsulating mRNA. LNPs containing beta-sitosterol showed superior ability to escape from endosomal entrapment compared to standard cholesterol LNPs. Live-cell imaging demonstrated a 10-fold increase in detectable endosomal perturbation events with LNPs containing beta-sitosterol.
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 a galectin 8-GFP (Gal8-GFP) cell reporter system to visualize the endosomal escape capabilities of LNP-encapsulated 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, 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 advancements in gene delivery.
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