Project description:Analysis of smallRNA in THP1 (human monocytic leukemia) cell line in order to correlate miRNA activity with target abundance. THP1 smallRNA profiles were generated in triplicates by deep-sequencing in Illumina HiSeq2000.
Project description:SmallRNAs are proposed as key regulators in many cellular processes including angiogenesis, suggested candidates for future therapeutic applications. But, regulation and modulation of smallRNAs in pathology conditions and normal conditions are poorly recognized, supremely in wound care management. Current study focused to identify the smallRNA regulatory network in simulated microgravity sensitized Human Umbilical Cord Vein Endothelial cells (HUVECs) and gravity (1g) as a background. HUVECs were purchased from Lonza (Cat.No) and cultured in EGM2 medium (Cat.No CC3162) supplemented with 10% Fetal Bovine, 1% penicillin-streptomycin (W/V). Cells were cultured in RPM (Randomized positioned machine) for 2 hours at 37°C.
Project description:We performed a smallRNA-Seq analysis comparing thymic lymphoma tissues from the p53-null (n=4) and ΔNp63Δ/Δ;p53-/- (n=3). Mice at 10 weeks of age were injected with either Ad-mCherry or Ad-CRE-mCherry to delete ΔNp63 in the thymic lmyphomas. We aimed to test by deleting the ΔNp63 in these p53-deficient tumors will mediate tumor regression and analyze the expression profile of the small RNAs
Project description:Purpose: The goals of this study is to compare and profile the smallRNA transcriptome of the placenta in preeclamptic and normal patients using RNA sequencing. Methods: Placental and Placental vesicles (STB-EVs) smallRNA profiles of normal and preeclamptic patients were generated by deep sequencing using Illumina HISEQ. FASTq.gz files were compressed with OASIS compressor and alignment was done with OASIS 2.0 ( by trimmimng with trimmomatic, aligning using default OASIS 2.0 aligning papameters). Quantitative PCR validation was performed using TaqMan gene expression assays Results: This contains a set of three parallel smallRNA sequencing experiments involving placenta tissue, medium/large STB-EVs and small STB-EVs. Comparison between PE and normal pregnancy placental tissue revealed 134 (p-value of <0.05 ) while in medium/large STB-EVs, 101 and in small STB-EVs, 16 (adjusted P-value of <0.05) differentially expressed small RNA We identified a number of mechanistic and biomarker targets, which were validated with qRT–PCR and confirmed to be signifficantly DE. The differentially expressed analysis identified potential yet undescribed small RNAs that may contribute to the pathogenesis of preeclampsia or/and may act as biomarkers of the disease. Conclusions: Our study represents the first combined analysis of placenta, medium/large and small STB-EV transcriptomes, with biologic replicates, generated by RNA-seq technology. The optimized data analysis workflows reported here should provide a framework for comparative investigations of expression profiles. Our results identified potential Placenta EV small RNA biomarkers that can help diagnose the preeclampsia
Project description:To identify microRNAs as TLR-activating molecules we have induced apoptosis in primary cortical neurons to trigger specific microRNA release into media. smallRNA sequencing revealed potential microRNAs ligands derived from dying neurons and their corresponding media that harbor already described TLR7/8 activating sequences. In line with this finding, about 80% of our differentially present miRNAs were able to activate TLR7/8 reporter cell lines. Furthermore, microRNAs miR-340-3p and miR-132-5p induced a specific cytokine and chemokine release form microglia leading to neuronal death. Within our systematic approach, we have identified TLR paralog-specific activating microRNAs with the potential to serve as biomarkers for brain diseases.
