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Endocytic Profiling of Cancer Cell Models Reveals Critical Factors Influencing LNP-Mediated mRNA Delivery and Protein Expression
How Cancer Cell Features Affect Lipid Nanoparticle Delivery of mRNA and Protein Production
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Abstract
A 30-cell line transfection screen revealed three cell lines with varying transfection efficiencies that correlated with protein expression.
- High-transfecting cells exhibited rapid uptake and trafficking of lipid nanoparticles through an organized pathway to lysosomes or exocytosis.
- Low-transfecting cells showed slower trafficking of lipid nanoparticles to lysosomes, alongside disorganized endocytic processes and poor acidification.
- Differences in endolysosomal morphology, localization, uptake, trafficking, recycling, and pH were identified among the cell lines using a novel pH probe.
- Efficient lipid nanoparticle-mRNA transfection may depend on a brief and specific time frame for endosomal escape before lysosomal degradation occurs.
- Endocytic profiling could be a key step in pre-clinical evaluations to optimize nucleic acid delivery systems for clinical applications.
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