Casein kinase II promotes target silencing by miRISC through direct phosphorylation of the DEAD-box RNA helicase CGH-1
Ontology highlight
ABSTRACT: In this study we show that global levels of mature miRNAs are unaffected in C. elegans after knockdown of either subunit of CK2 (encoded by kin-3 and kin-10) by RNAi. Small RNAs were quantified from wild-type animals at L4 stage fed either vector (L4440), kin-3, kin-10, or alg-1 RNAi. Mature miRNA counts were quantified by summing the total number of reads mapping to each miRNA and normalized to the total number of mapped reads per sequencing library.
Project description:SPF leghorn chickens were infected with C. jejuni. The cecum were collected at 8h post infection for total RNA isolation. The significantly expressed microRNAs between infected and non-infected chickens were identified through Solexa sequencing technology. Two libraries were made from each group, in total, four libraries were used for Solexa sequencing.
Project description:To determine the effect of Egr2 and Egr3 on tumour infiltrating lymphocyte gene expression, GFP-Egr2 knockin (Egr2 Kin) and hCD2-Cre Egr2loxP/loxP Egr3-/- (Egr2/3 DKO) mice were inoculated with MC38 or B16 tumour cells. Fourteen days later, GFP-Egr2high and Egr2-/-Egr3-/- CD8+ tumour infiltrating lymphocytes were isolated from Egr2 Kin and Egr2/3 DKO mice, respectively, and analysed by RNA-seq.
Project description:Transcription profiling of naive and memory phenotype mouse CD4+ T cells extracted from GFP-Egr2 knockin (Egr2 Kin) and Egr2loxP/loxP hCD2-Cre Egr3-/- (Egr2/3 DKO) mice in the steady state
Project description:Transcription profiling of mouse CD4+ and CD8+ T cells extracted from GFP-Egr2 knockin (Egr2 Kin) and hCD2-Cre / Egr2loxP/loxP / Egr3-/- Egr2/3 DKO) mice 7 days after infection with vaccinia virus.
Project description:Transcription profiling of naive and memory phenotype mouse CD8+ T cells extracted from GFP-Egr2 knockin (Egr2 Kin) and Egr2loxP/loxP hCD2-Cre Egr3-/- (Egr2/3 DKO) mice in the steady state
Project description:In nature, animals often face feast or famine conditions. We aimed to identify the miRNAs of Caenorhabditis elegans that changed their expression under starvation conditions in stage L4 larvae. Our results highlight 14 miRNAs that show differential expression in starved versus well-fed larvae. In particular, miRNAs of the miR-35-3p/miR-41-3p family were upregulated 6-20 fold upon starvation. We verified the upregulation of miR-35-3p with qPCR. Additionally, we showed that the expression of gld-1, important in ovogenesis, and a validated target of miR-35-3p, was downregulated when the expression of miR-35-3p was higher. This study represents a starting point for a more comprehensive understanding of the role of miRNAs during starvation in C. elegans. Illumina small RNA sequencing of starved and well-fed L4 worms.
Project description:Transcription profiling of mouse FR4+Egr2+, FR4+Egr2- and FR4-Egr2- CD4+ T cells extracted from GFP-Egr2 knockin (Egr2 Kin) and FR4+Egr2-/-Egr3-/- and FR4-Egr2-/-Egr3-/- CD4+ T cells isolated from Egr2loxP/loxP hCD2-Cre Egr3-/- (Egr2/3 DKO) mice in the steady state or 21 days after infection with vaccinia virus.
Project description:Sequencing of the 3â end of poly(A)+ RNA identifies cleavage and polyadenylation sites (pAs) and measures transcript expression. We previously developed a method, 3â region extraction and deep sequencing (3âREADS), to address mispriming issues that often plague 3â end sequencing. Here we report a new version, named 3âREADS+, which has vastly improved accuracy and sensitivity. Using a special locked nucleic acid oligo to capture poly(A)+ RNA and to remove bulk of the poly(A) tail, 3âREADS+ generates RNA fragments with an optimal number of terminal As that balance data quality and detection of genuine pAs. With improved RNA ligation steps for efficiency, the method shows much higher sensitivity (over two orders of magnitude) compared to the previous version. Using 3âREADS+, we have uncovered a sizable fraction of previously overlooked pAs located next to or within a stretch of adenylate residues in human genes, and more accurately assessed the frequency of alternative cleavage and polyadenylation (APA) in HeLa cells (~50%). 3âREADS+ will be a useful tool to accurately study APA and to analyze gene expression by 3â end counting, especially when the amount of input total RNA is limited. Nine 3'READS+ libraries were made with different amounts (100 ng, 200 ng, 400 ng, 1000 ng, 5000 ng, 15000 ng) of input Hela RNA.
Project description:Porcine cytomegalovirus (PCMV; genus Cytomegalovirus, subfamily Betaherpesvirinae, family Herpesviridae) is an immunosuppressive virus that mainly inhibits the immune function of T lymphocytes and macrophages, which has caused great distress to the farming industry. In this study, we obtained the miRNA expression profiles of PCMV-infected and control porcine macrophages, PCMV-infected and control porcine tissues via high-throughput sequencing. The comprehensive analysis of miRNA profiles showed that 306 miRNA database annotated and 295 novel pig-encoded miRNAs were detected. Gene Ontology (GO) analysis of the target genes of miRNAs in PCMV infected porcine macrophages showed that the differentially expressed miRNAs are mainly involved in immune and metabolic process. This is the first report of the miRNA transcriptome in PCMV infected porcine macrophages and PCMV infected tissues and the analysis of the miRNA regulatory mechanism during PCMV infection. Further research into the regulatory mechanisms of miRNAs during immunosuppressive viral infections will contribute to the treatment and prevention of immunosuppressive viruses. miRNA expression profiling of PCMV-infected and control porcine macrophages; PCMV-infected and control porcine tissues via high-throughput sequencing.
Project description:To explore the differences in miRNA profiles, naive CD4+CD62L+ helper T cells were sorted by magnetic cell sorting from spleens of female C57BL/6 mice and were induced to differentiate into Th1 or Th17 cells, then total RNA was extracted and miRNA-seq were conducted. The results showed that many microTNAs presented a different expression pattern between Th1 and Th17 cells, which prompted us to further study their roles in Th117 differenciation. miRNA profiles of Th0 and Th17 cells were generated by deep sequencing, in triplicate, using Illumina HiSeq 2500