Project description:We report the gene expression in L1 arrest (48 hours) of wild-type sleeping worms (N2) and RIS-manipulated worms (HBR2340 and PHX2193)
Project description:We used RNA-seq analysis to identify changes in gene expression occurring in linc-133 (aqz 3) mutants comparing with that in wild type (N2) worms during L1 arrest stage. Basing on these sequencing data, genes which were up-regulated or down-regulated by at least 2 times( |log2(FoldChange)| ≥ 1 and padj ≤ 0.05) subsequent to WT worms were analyzed. These differentially expressed genes were analyzed for pathway and gene ontology (GO) enrichment, which indicated that the genes influenced by aqz 3 mutation of linc-133 gene were enriched in several significant signaling pathways, including longevity, protein degradation in endoplasmic reticulum pathway, and fat metabolism pathways.
Project description:Transcriptional profiling of N2 vs. mir-243 worms, aiming to identify direct and indirect targets of the microRNA. N2 and mir-243 young-adult worms grown at 20C were analyzed. Three biological replicates with dye-swaps were performed.
Project description:Transcription factors are key components of regulatory networks that control development, as well as the response to environmental stimuli. We have established an experimental pipeline in C. elegans that permits global identification of the binding sites for transcription factors using chromatin immunoprecipitation and deep sequencing. We describe and validate this strategy, and apply it to the transcription factor PHA-4, which plays critical roles in organ development and starvation stress response. We defined binding sites for PHA-4 during formation of the embryonic pharynx, and in starved larvae. PHA-4 binds to thousands of locations throughout the genome. These sites shift dramatically between embryos and larvae, from developmentally regulated genes to genes involved in metabolism. We acquired transcriptome of wild type N2 worms at embyonic and starved L1 stages using RNA-seq method in comparing to the ChIP-seq experiment to study transcription factor binding. The wild type worms were cultured the same as the transgenic worms for transcription factor binding experiment. Keywords: Transcriptome Analysis For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf Two development stages embryo and starved larvae each contains 3 technical replicates are reported
Project description:Transcription factors are key components of regulatory networks that control development, as well as the response to environmental stimuli. We have established an experimental pipeline in C. elegans that permits global identification of the binding sites for transcription factors using chromatin immunoprecipitation and deep sequencing. We describe and validate this strategy, and apply it to the transcription factor PHA-4, which plays critical roles in organ development and starvation stress response. We defined binding sites for PHA-4 during formation of the embryonic pharynx, and in starved larvae. PHA-4 binds to thousands of locations throughout the genome. These sites shift dramatically between embryos and larvae, from developmentally regulated genes to genes involved in metabolism. We acquired transcriptome of wild type N2 worms at embyonic and starved L1 stages using RNA-seq method in comparing to the ChIP-seq experiment to study transcription factor binding. The wild type worms were cultured the same as the transgenic worms for transcription factor binding experiment. Keywords: Transcriptome Analysis For data usage terms and conditions, please refer to http://www.genome.gov/27528022 and http://www.genome.gov/Pages/Research/ENCODE/ENCODEDataReleasePolicyFinal2008.pdf
Project description:Eggs were isolated by hypochlorite treatment and allowed to hatch in the absence of food as L1 stage arrested worms in S-buffer. Worms were then transferred to large plates containing NGM alone or NGM+ resveratrol and allowed to grow till young adult stage (ca 40 hours). Worms were harvested and total RNA extracted using Trizol reagent. Poly-A+ RNA was isolated using Qiagen midi-prep kit and used for microarray hybridization. A compound treatment design type is where the response to administration of a compound or chemical (including biological compounds such as hormones) is assayed. Compound Based Treatment: Resveratrol Computed
Project description:Purpose: Comparing the transcriptome of wild-type(N2), hlh-11 knock-out and hlh-11 over-expression worms to identify genes with differential expression. Methods: For each condition, two replicates were examined. For each sample, approximately 2,000 synchronized young adult worms were collected and resuspended in TRIzon Reagent (CW0580, CWBIO). Total RNA was isolated by chloroform extraction, followed by isopropanol precipitation. Sequencing was performed using HiSeq2500. The clean reads were aligned to WBcel235 genomes with TopHat2. Gene differential expression was analyzed using DEseq2. A threshold of p < 0.05 was set to determine differentially expressed genes. Induction (KO_up or OE_up) or suppression (KO_down or OE_down) of genes were obtained through comparing gene expressions in hlh-11 KO or OE worms to wild-type N2. Results: >86% reads were uniquely mapped. 2637 genes were significantly up regulated and 2845 genes were significantly down regulated in hlh-11 knock-out worms compared to wild-type. 107 genes were significantly up regulated and 237 genes were significantly down regulated in hlh-11 over-expression worms compared to wild-type. Conclusions: This study revealed the genes transcriptionally regulated by hlh-11.