Unravelling cytosolic delivery of cell penetrating peptides with a quantitative endosomal escape assay
| Autor: | Angus P. R. Johnston, Daniel Yuen, Colin W. Pouton, Joshua J. Rennick, Serena L. Y. Teo, Hareth Al-Wassiti |
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| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
Endosome Cell Survival Science Cell Green Fluorescent Proteins General Physics and Astronomy Gene Expression 02 engineering and technology Cell-Penetrating Peptides Endosomes Sensitivity and Specificity General Biochemistry Genetics and Molecular Biology Article Mass Spectrometry Cell membrane 03 medical and health sciences Cytosol Cell Line Tumor medicine Humans Cytosolic transport Luciferase Luciferases Sensors and probes Multidisciplinary Protein transport Chemistry Protein delivery Cell Membrane General Chemistry 021001 nanoscience & nanotechnology Recombinant Proteins Cell biology Transport protein 030104 developmental biology medicine.anatomical_structure Luminescent Measurements Nucleic acid 0210 nano-technology |
| Zdroj: | Nature Communications, Vol 12, Iss 1, Pp 1-12 (2021) Nature Communications |
| ISSN: | 2041-1723 |
| Popis: | Cytosolic transport is an essential requirement but a major obstacle to efficient delivery of therapeutic peptides, proteins and nucleic acids. Current understanding of cytosolic delivery mechanisms remains limited due to a significant number of conflicting reports, which are compounded by low sensitivity and indirect assays. To resolve this, we develop a highly sensitive Split Luciferase Endosomal Escape Quantification (SLEEQ) assay to probe mechanisms of cytosolic delivery. We apply SLEEQ to evaluate the cytosolic delivery of a range of widely studied cell-penetrating peptides (CPPs) fused to a model protein. We demonstrate that positively charged CPPs enhance cytosolic delivery as a result of increased non-specific cell membrane association, rather than increased endosomal escape efficiency. These findings transform our current understanding of how CPPs increase cytosolic delivery. SLEEQ is a powerful tool that addresses fundamental questions in intracellular drug delivery and will significantly improve the way materials are engineered to increase therapeutic delivery to the cytosol. Our understanding of cytosolic delivery is hindered by existing methods for quantification which suffer from being indirect and showing low sensitivity. Here the authors report a SLEEQ (Split Luciferase Endosomal Escape Quantification) assay to assess cytosolic delivery of cell-penetrating peptides. |
| Databáze: | OpenAIRE |
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