Project description:Communication between the maternal uterus and the embryo is vital for a successful pregnancy. Exosomes, subtypes of extracellular vesicles comprising many bioactive factors regulate the early stages of pregnancy, specifically during embryo implantation. Nevertheless, the mechanism by which exosomal microRNAs (miRNAs) derived from placental trophoblasts regulate embryo implantation remains elusive. Herer, we isolated and identified exosomes derived from placental trophoblasts cells (HTR8/SVneo). Subsequently, we evaluated the loading miRNA in exosomes by small RNA sequencing. This study provides novel insights into the mechanism of trophoblasts cells-derived exosomes during embryo implantation.
Project description:We have sequenced miRNA libraries from human embryonic, neural and foetal mesenchymal stem cells. We report that the majority of miRNA genes encode mature isomers that vary in size by one or more bases at the 3’ and/or 5’ end of the miRNA. Northern blotting for individual miRNAs showed that the proportions of isomiRs expressed by a single miRNA gene often differ between cell and tissue types. IsomiRs were readily co-immunoprecipitated with Argonaute proteins in vivo and were active in luciferase assays, indicating that they are functional. Bioinformatics analysis predicts substantial differences in targeting between miRNAs with minor 5’ differences and in support of this we report that a 5’ isomiR-9-1 gained the ability to inhibit the expression of DNMT3B and NCAM2 but lost the ability to inhibit CDH1 in vitro. This result was confirmed by the use of isomiR-specific sponges. Our analysis of the miRGator database indicates that a small percentage of human miRNA genes express isomiRs as the dominant transcript in certain cell types and analysis of miRBase shows that 5’ isomiRs have replaced canonical miRNAs many times during evolution. This strongly indicates that isomiRs are of functional importance and have contributed to the evolution of miRNA genes
Project description:Matrix-bound extracellular vesicles were isolated from the left ventricle tissue of de-identified patients with non-failing (healthy) hearts or with non-ischemi heart failure. mIRNA sequencing was performed on the miRNA cargo sequestered within the extracellular vesicles. Differences in the miRNA signature of healthy versus failing-tissue derived extracellular vesicles suggests that disease progression to heart failure is associated with dynamic changes in vesicular cargo.
Project description:miRNA-sequencing of grapefruit-derived extracellular vesicles and fusion nanovesicles derived from grapefruit-derived extracellular vesicles and gingival mesenchymal stem cell-derived vesicles. We then performed gene expression profiling analysis to explore the miRNAs derived from grapefruit-derived extracellular vesicles, and the retention rate of miRNAs after membrane fusion
Project description:To gain insight into the microRNA expression profile of small extracellular vesicles derived from bone metabolism related cell types and to verify their mechanism, we utilized the miRNA sequencing technology to analyze the miRNA profiles of different mouse osteoblast and osteoclast cell derived small extracellular vesicles.
2022-05-26 | GSE203618 | GEO
Project description:miRNA Sequencing Small Extracellular Vesicles
Project description:We report the differential abundance of cell free miRNAs extracted from extracellular vesicles (EV) in the peripheral blood of pregnant women with or without preeclampsia by Next Generation Sequencing. Maternal blood samples were collected form pregnant women enrolled in the study during first to very early second trimester (11–14 weeks ), mid- to late second trimester (19–22 weeks), third trimester (36 weeks) and at delivery. Extracellular vesicles were isolated from peripheral blood and total RNA was extracted using the miRNeasy Mini Kit (Qiagen, Valencia, CA) following manufacturer’s instruction. miRNA libraries were prepared utilizing NEB Next Multiplex small RNA Library prep kit (NEB E7300S; New England Biolabs, Inc, Ipswich, MA, USA) and libraries were subsequently sequenced using the HiSeq-2500 platform with single-end 50bp reads (Illumina Inc.; San Diego, CA, USA).
