Applications of cell resealing to reconstitute microRNA loading to extracellular vesicles
| Autor: | Masayuki Murata, Fumi Kano, Yuki Sonoda |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Endosome
Science Cytological Techniques Cell RNA-binding proteins Membrane trafficking Biochemistry Membrane Fusion Article Exocytosis Fluorescence imaging Extracellular Vesicles medicine Extracellular Humans Secretion Super-resolution microscopy Analytical biochemistry Multivesicular bodies Multidisciplinary Chemistry Binding protein Argonaute Endocytosis Cell biology Confocal microscopy MicroRNAs Cytosol medicine.anatomical_structure miRNAs MiRNA transport Nanoparticles Medicine HeLa Cells |
| Zdroj: | Scientific Reports, Vol 11, Iss 1, Pp 1-18 (2021) Scientific Reports |
| Popis: | MicroRNAs (miRNAs) are cargo carried by extracellular vesicles (EVs) and are associated with cell–cell interactions. The response to the cellular environment, such as disease states, genetic/metabolic changes, or differences in cell type, highly regulates cargo sorting to EVs. However, morphological features during EV formation and secretion involving miRNA loading are unknown. This study developed a new method of EV loading using cell resealing and reconstituted the elementary miRNA-loading processes. Morphology, secretory response, and cellular uptake ability of EVs obtained from intact and resealed HeLa cells were comparable. Exogenously added soluble factors were introduced into multivesicular endosomes (MVEs) and their subsequent secretion to the extracellular region occurred in resealed HeLa cells. In addition, miRNA transport to MVEs and miRNA encapsulation to EVs followed a distinct pathway regulated by RNA-binding proteins, such as Argonaute and Y-box binding protein 1, depending on miRNA types. Our cell-resealing system can analyze disease-specific EVs derived from disease model cells, where pathological cytosol is introduced into cells. Thus, EV formation in resealed cells can be used not only to create a reconstitution system to give mechanistic insight into EV encapsulation but also for applications such as loading various molecules into EVs and identifying disease-specific EV markers. |
| Databáze: | OpenAIRE |
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