Polyethylene Glycol -Mediated Exosome Isolation: A Method for Exosomal RNA Analysis.
Exosomes
Extracellular vesicles
MicroRNA
Journal
Iranian biomedical journal
ISSN: 2008-823X
Titre abrégé: Iran Biomed J
Pays: Iran
ID NLM: 9814853
Informations de publication
Date de publication:
24 Feb 2024
24 Feb 2024
Historique:
medline:
12
3
2024
pubmed:
12
3
2024
entrez:
12
3
2024
Statut:
aheadofprint
Résumé
ExoRNAs offer valuable insights into their cellular origin. ExoRNA studies were faced with challenges in obtaining sufficient amounts of high-quality RNA. Herein, we aimed to compare three traditional exosome isolation methods to introduce an appropriate strategy to extract RNA from cancer-derived exosomes for further RNA analysis. Exosomes were isolated through ultracentrifugation, precipitation kit, and size exclusion column chromatography, and then characterized by DLS and TEM, followed by extracting total RNA. The quality and quantity of the extracted RNAs were assessed by a NanoDrop and 2.5% agarose gel electrophoresis. Extracted exosomes displayed a similar range of size and morphology. We found that PEG-precipitation method resulted in a higher RNA yield with a 260/280 ratio of 1.9. The obtained exoRNA appeared as a smear in the agarose gel, indicative of small exoRNAs. We provide researchers a suitable approach to isolate exosomes based on yield and purity of exoRNA.
Sections du résumé
Background
UNASSIGNED
ExoRNAs offer valuable insights into their cellular origin. ExoRNA studies were faced with challenges in obtaining sufficient amounts of high-quality RNA. Herein, we aimed to compare three traditional exosome isolation methods to introduce an appropriate strategy to extract RNA from cancer-derived exosomes for further RNA analysis.
Methods
UNASSIGNED
Exosomes were isolated through ultracentrifugation, precipitation kit, and size exclusion column chromatography, and then characterized by DLS and TEM, followed by extracting total RNA. The quality and quantity of the extracted RNAs were assessed by a NanoDrop and 2.5% agarose gel electrophoresis.
Results
UNASSIGNED
Extracted exosomes displayed a similar range of size and morphology. We found that PEG-precipitation method resulted in a higher RNA yield with a 260/280 ratio of 1.9. The obtained exoRNA appeared as a smear in the agarose gel, indicative of small exoRNAs.
Conclusion
UNASSIGNED
We provide researchers a suitable approach to isolate exosomes based on yield and purity of exoRNA.
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM