Project description:EV RNA samples from MH-S cells were prepared for small RNA sequencing by TruSeq Small RNA Sample Prep Kits (Illumina, San Diego，USA), using a minimum of 1 μg RNA per sample.
Project description:We report the high-throughput profiling of small RNA in extracellular vesicles (EVs) and their producing mammalian cells using Illumina small RNA sequencing platform. By obtaining over 2190 miRNA expression from 18 samples, the miRNA profile in EVs and their producing cells were compared. Results provide insight into gene profile difference between EV loading and the producing cells.
Project description:As a part of study to characterize the effects of fluid flow shear stress to mouse muscle cells, small RNA sequencing was performed with muscle cell-derived extracellular vesicles (Myo-EVs).
Project description:Thy1 + cells isolated from male F344 rats with Dgalactosamine-induced liver injury were cultured for one week. Bone marrow mesenchymal cells isolated from from the femurs and tibias of a 5-week old male F344 rat. Small hepatocytes (SH) and mature hepatocytes (MH) were isolated from normal livers of 7-week old male F344 rat, Extracellular vesicles (EVs) were separedt from culture media by ultracentrifugation.
Project description:Understanding the functional role of RNA modifications and to what extent they might regulate mature miRNA function and their secretion in extracellular vesicles (EVs) is unknown. Analysis of RNA content across a variety of EVs has shown differential miRNA enrichment when compared to parental cell expression patterns. Whether this is due to differential stability, specific regulation of miRNA export, or other mechanisms is unclear. Here, we tested whether RNA base modifications and recognition of those modifications might underlie differential miRNA export into EVs derived from KRAS mutant cell lines. We found that decreased levels of Mettl3, a writer of N6-methyladenosine (m6A) modification, altered extracellular transfer of miRNAs containing consensus sequences for m6A. Further, EVs prepared from cells expressing shRNAs against Mettl3 were incapable of conferring colony growth in 3D to wild type KRAS cells. Our data indicate that m6A modification plays an important role in miRNA export to EVs.
