Hypothalamic DNA-methylation differences in Red Junglefowl divergently selected for reduced fear of humans
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ABSTRACT: Domestication of animals causes large phenotypic alterations during a brief evolutionary window of time. These alterations are caused by genomic variation, yet the prevalence of modified traits is larger than expected if caused only by classical genetics and mutations. Two selection lines of Red Junglefowl (ancestors of modern chickens), bred for either high or low fear of human for five generations, were used to study the differences in hypothalamic DNA methylation between the two populations.
Project description:Methylated DNA immunoprecipitation sequencing (MeDIP-Seq) is a widely used approach to study DNA methylation genome-wide. Here, we present a novel MeDIP-Seq protocol compatible with the Ion Torrent semiconductor-based sequencing platform that is scalable and accurately identifies sites of differential DNA methylation. Additionally, we demonstrate that the high-throughput data derived from MeDIP-Seq on the Ion Torrent platform provides adequate coverage of CpG cytosines, the methylation states of which we validated at single-base resolution on the Infinium HumanMethylation450K Beadchip array. We applied this integrative approach to further investigate the role of DNA methylation in alternative splicing and to profile 5-mC and 5-hmC variants of DNA methylation in normal human brain tissue that we observed localize over distinct genomic regions. These applications of MeDIP-Seq on the Ion Torrent platform have broad utility and add to the current methodologies for profiling genome-wide DNA methylation states in normal and disease conditions. MeDIP-Seq on Ion Torrent Platform in HCT116 and Human Brain
Project description:We investigated a panel of 21 genes by parallel sequencing on the Ion Torrent Personal Genome Machine platform. We sequenced 65 CRCs that were treated with cetuximab or panitumumab ( 37 samples were responsive and 28 were resistant).
Project description:TGFB2-AS1 is a long non-coding RNA which is induced by ΤGFβ signaling. In order to assess the importance of TGFB2-AS1 on the regulation of gene expression, we performed an AmpliSeq transcriptomic array in human keratinocytes (HaCaT), which stably over-express TGFB2-AS1 or control pcDNA3 empty vector. In addition, cells were stimulated with TGFβ1 for 24 hours, in order to observe the effects of TGFB2-AS1 on gene expression, downstream of TGFβ signaling. RNA from the following four conditions was used in this experiment: 1) pcDNA3, 2) pcDNA3+TGFβ1, 3) pcDNA3-TGFB2-AS1, 4) pcDNA3-TGFB2-AS1+TGFβ1. Biological triplicates were used per condition.
Project description:Estrogens receptor a (ERα) is essential for breast tumors,since about seventy percent of breast cancers are detected as ERα positive.Recent studies suggest that ERα is related with the epithelial cell morphology. Recently, it has demonstrated that the suppression of ERα induced epithelial-mesenchymal transition (EMT) in the MCF-7 breast cacner cells. Interestingly, the loss of ERa resulted in strong differences on the gene expression profile of a variety of genes. Therefore, the aim of the RNA-seq is to elucidate the effect of the silencing of ERα on the mRNA levels of a larger variety of genes, thus revealing possible target genes which may be implicated on the aggressive phenotype and behavior of the ERα-suppresed MCF-7/SP10+ breast cancer cells. For this reason total RNA from both MCF-7/SP10+ cells and of their internal control MCF-7/C cells was extracted in 3 biological replicates and 3 technical replicates.
Project description:Molecular profiling of breast cancer has achieved great depth in defining the mutational landscapes and molecular profiles of primary tumors, though little is still known regarding cancer evolution into a recurrence. Proteogenomic workflows are particularly useful in defining the multi-layered biology of complex diseases by combining genomic, transcriptomic, and proteomic technologies so to inform not only on mutational processes, but also on their repercussion at the effector level, proteins. We employed our recently developed proteogenomic workflow to analyze a cohort of 27 primary breast cancers and their matched loco-regional recurrences by whole genome sequencing, RNA sequencing, and mass spectrometry.
Project description:Stomata pores are surrounded by a pair of guard cells in the epidermis in plants. Stomata apature is controlled by environmental factors. Stomata opening is also enhanced by introduction of SOC1-GFP driven by GC1 promoter in phot1 phot2 double mutants (pGC1::SOC1-GFP/ phot1 phot2). Epidermis including guard cells were isolated from phot1 phot2 and pGC1::SOC1-GFP/phot1 phot2 and gene expression were analyzed. phot1-5 phot2-1 in gl1 background (phot1 phot2) and pGC1::SOC1-GFP/phot1 phot2 were grown under 16 h light / 8h dark, constant 22?C conditions for 4 to 5 weeks. Epidermis including guard cells were isolated from leaves of these plants. Three biological replicates were used.
Project description:Obesity is well recognized as a risk factor for coronary heart disease and mortality. The relationship between abdominal obesity and ischemic stroke remains less clear. Previous publication showed the obesity is an independent, potent risk factor for ischemic stroke in all race-ethnic groups. It is a stronger risk factor than BMI and has a greater effect among younger persons. The goal of this experiment was to compare genome wide enrichment of H3K9ac histone mark profile of white blood cells of healthy controls, patients with obesity and/or stroke in order to understand the histone modifications differences behind the different phenotypes. There were 3 subjects in each group.
Project description:Purpose: The aim of this study is to investigate the translational regulation of skeletal muscle during acute endurance exercise. Methods: We used mRNA-Seq and ribosome profiling to examine transcriptional and translational regulation, respectively. Result: There were clear distinctions between the profiles of transcription and translation even at a basal condition. TOP-motif genes were translationally suppressed immediately after the exercise. Other genes, such as Slc25a25 was significantly translationally up-regulated presumably in a mTOR-independent manner. Conclusion: There were diverse regulation between transcription and translation. Although many focused on overall protein synthesis to understand the effect of exercise, translational regulation of individual genes are required. Transcriptional and translational profiles of mouse gastrocnemius with or without acute endurance exercise were generated using Ion PGM sequencer.