Isolation of Exosome-Like Nanoparticles and Analysis of MicroRNAs Derived from Coconut Water Based on Small RNA High-Throughput Sequencing

• Published in: Journal of agricultural and food chemistry 2018-03, Vol.66 (11), p.2749-2757
• Main Authors: Zhao, Zhehao, Yu, Siran, Li, Min, Gui, Xin, Li, Ping
• Format: Article
• Language: English
• Subjects: Cocos - chemistry; Cocos - classification; Cocos - genetics; Cocos - metabolism; Endosperm - genetics; Endosperm - growth & development; Exosomes - chemistry; Exosomes - genetics; Exosomes - metabolism; High-Throughput Nucleotide Sequencing; Humans; MicroRNAs - genetics; MicroRNAs - metabolism; Nanoparticles - chemistry; Polymerase Chain Reaction; Real-Time Polymerase Chain Reaction; RNA, Plant - genetics; RNA, Plant - metabolism
• ISSN: 0021-8561; 1520-5118
• DOI: 10.1021/acs.jafc.7b05614

In this study, the presence of microRNAs in coconut water was identified by real-time polymerase chain reaction (PCR) based on the results of high-throughput small RNA sequencing. In addition, the differences in microRNA content between immature and mature coconut water were compared. A total of 47 known microRNAs belonging to 25 families and 14 new microRNAs were identified in coconut endosperm. Through analysis using a target gene prediction software, potential microRNA target genes were identified in the human genome. Real-time PCR showed that the level of most microRNAs was higher in mature coconut water than in immature coconut water. Then, exosome-like nanoparticles were isolated from coconut water. After ultracentrifugation, some particle structures were seen in coconut water samples using 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanine perchlorate fluorescence staining. Subsequent scanning electron microscopy observation and dynamic light scattering analysis also revealed some exosome-like nanoparticles in coconut water, and the mean diameters of the particles detected by the two methods were 13.16 and 59.72 nm, respectively. In conclusion, there are extracellular microRNAs in coconut water, and their levels are higher in mature coconut water than in immature coconut water. Some exosome-like nanoparticles were isolated from coconut water, and the diameter of these particles was smaller than that of animal-derived exosomes.