Enhanced Loading of Functional miRNA Cargo via pH Gradient Modification of Extracellular Vesicles
| Autor: | Wei Chao, Eshan Dahal, Stephanie M. Kronstadt, Daniel Levy, Lin Zou, Daphne R Knudsen, Steven M. Jay, Sheng Wang, Babita Parajuli, Anjana Jeyaram, Tek N. Lamichhane |
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| Rok vydání: | 2020 |
| Předmět: |
Small interfering RNA
Extracellular Vesicles 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Nucleic Acids Drug Discovery microRNA Genetics Humans Molecular Biology 030304 developmental biology Pharmacology 0303 health sciences Electroporation Vesicle RNA Biological Transport Hydrogen-Ion Concentration Microvesicles MicroRNAs HEK293 Cells chemistry 030220 oncology & carcinogenesis Nucleic acid Biophysics Molecular Medicine Original Article DNA |
| Zdroj: | Mol Ther |
| ISSN: | 1525-0016 |
| Popis: | Based on their identification as physiological nucleic acid carriers in humans and other organisms, extracellular vesicles (EVs) have been explored as therapeutic delivery vehicles for DNA, RNA, and other cargo. However, efficient loading and functional delivery of nucleic acids remain a challenge, largely because of potential sources of degradation and aggregation. Here, we report that protonation of EVs to generate a pH gradient across EV membranes can be utilized to enhance vesicle loading of nucleic acid cargo, specifically microRNA (miRNA), small interfering RNA (siRNA), and single-stranded DNA (ssDNA). The loading process did not impair cellular uptake of EVs, nor did it promote any significant EV-induced toxicity response in mice. Cargo functionality was verified by loading HEK293T EVs with either pro- or anti-inflammatory miRNAs and observing the effective regulation of corresponding cellular cytokine levels. Critically, this loading increase is comparable with what can be accomplished by methods such as sonication and electroporation, and is achievable without the introduction of energy associated with these methods that can potentially damage labile nucleic acid cargo. |
| Databáze: | OpenAIRE |
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