Project description:X-linked Dystonia-Parkinsonism (XDP) is a long-standing quandary in human disease genetics. XDP is predominantly observed on Panay island in the Philippines. This study is one of the first of its kind to interrogate an unsolved Mendelian disorder by integrating genome and transcriptome assembly methods using Illumina, 10X Genomics, Pacific Biosciences, and Agilent genome targeting technologies. These data provide strong evidence for a pathogenic link between a noncoding SVA retrotransposon and XDP. We demonstrate that this Mendelian disorder is associated with a sine-VNTR-Alu (SVA) retrotransposon that inserted into the TAF1 gene and is shared by all XDP probands, yet never observed in controls from worldwide populations. Transcriptome assembly in iPSC-derived neural stem cells (NSCs) and neurons revealed that this SVA caused aberrant splicing and significant intron retention, which was negatively correlated with TAF1 expression. Remarkably, CRISPR/Cas9... (for more see dbGaP study page.)
Project description:In this study, we aim to present a global transcriptome analysis of medicinal/spice plant, Crocus sativus. We generated about 206 million high-quality reads from five tissues (corm, leaf, Tepal, stamen and stigma) using Illumina platform. We performed an optimized de novo assembly of the reads and estimated transcript abundance in different tissue samples. The transcriptome dynamics was studied by differential gene expression analyses among tissue samples. Overall design: We collected different tissue samples from the mature plants. Total RNA isolated from these tissue samples was subjected to Illumina sequencing. The sequence data was further filtered using NGS QC Toolkit to obtain high-quality reads. The filtered reads were used for de novo assembly optimization. The reads were further mapped to the saffron transcripts via CLC Genomics Workbench and differential gene expression analysis was performed using DESeq software.
Project description:In this study, we aim to present a global view of transcriptome dynamics in different rice cultivars (IR64, Nagina 22 and Pokkali) under control and stress conditions. More than 50 million high quality reads were obtained for each tissue sample using Illumina platform. Reference-based assembly was performed for each rice cultivar. The transcriptome dynamics was studied by differential gene expression analyses between stress treatment and control sample. We collected seedlings of three rice cultivars subjected to control (kept in water), desiccation (transferred on folds of tissue paper) and salinity (transferred to beaker containing 200 mM NaCl solution) treatments. Total RNA isolated from these tissue samples was subjected to Illumina sequencing. The sequence data was further filtered using NGS QC Toolkit to obtain high-quality reads. The filtered reads were mapped to Japonica reference genome using Tophat software. Cufflinks was used for reference-based assembly and differential gene expression was studied using cuffdiff software. The differentially expressed genes during various abiotic stress conditions were identified.
Project description:Crassulacean acid metabolism (CAM) is a water-use efficient adaptation of photosynthesis that has evolved independently many times in diverse lineages of flowering plants. We hypothesize that convergent evolution of protein sequence and temporal gene expression underpins the independent emergences of CAM from C3 photosynthesis. To test this hypothesis, we generated a de novo genome assembly and genome-wide transcript expression data for Kalanchoe fedtschenkoi, an obligate CAM species within the core eudicots with a relatively small genome (~260 Mb). Our comparative analyses identified signatures of convergence in protein sequence and re-scheduling of diel transcript expression of genes involved in nocturnal CO2 fixation, stomatal movement, heat tolerance, circadian clock and carbohydrate metabolism in K. fedtschenkoi and other CAM species in comparison with non-CAM species. These findings provide new insights into molecular convergence and building blocks of CAM and will facilitate CAM-into-C3 photosynthesis engineering to enhance water-use efficiency in crops.
Project description:mTOR inhibitor rapamycin is a well-known anticancer and immunosuppressant agent. Effects of rapamycin on zebrafish cells have not been previously studied using transcriptome analyses. Our microarray analysis on ZF4 cells showed that rapamycin treatment modulated a large set of genes with varying functions including protein synthesis, assembly of mitochondrial and proteosomal machinery, cell cycle, metabolism, and oxidative phosphorylation. Overall design: Zebrafish ZF4 cells exposed to 100nM rapamycin was profiled using Zebrafish Affymetrix Gene Chips. Normalization by RMA and class comparison analyses were performed using BRB Array Tools.
Project description:Our aim was to classify and quantify transcripts in primary duck hepatocytes cultured in medium with 5% FBS or 1.5% DMSO for 8 days. Methods: The transcriptome of PDHs under different conditions was analyzed by the pair-end sequencing on the Illumina Solexa platform. High-quality reads were mapped to the Anas platyrhynchos genome with TopHat v2.0.12 software. TopHat allows multiple alignments per read and default parameters were used. Cufflinks v2.2.1 software was later used for analyses that included transcript assembly and FPKM value calculations to quantify gene expression; this program was also run with default parameters. The transcriptome of PDHs under different culture conditions were analyzed by the paired-end sequencing on the Illumina Solexa platform.
Project description:DDX46 is identified to be required at the early step of pre-spliceosome assembly,but whether DDX46 could regulate antiviral transcripts isoform splicing in the nucleus remain elusive. Overall design: Transcriptome sequencing analyses of WT and DDX46+/- macrophages from mouse peritoneal macrophages infected with VSV for 8 h.
Project description:Colon cancer onset and progression is strongly associated with the presence, absence, or relative abundances of certain microbial taxa in the gastrointestinal tract. However, specific mechanisms affecting disease susceptibility related to complex commensal bacterial mixtures are poorly understood. We used a multi-omics approach to determine how differences in the complex gut microbiome (GM) influence the metabolome and host transcriptome and ultimately affect susceptibility to adenoma development. Fecal samples collected from a preclinical rat model of colon cancer harboring distinct complex GMs were analyzed using ultra-high performance liquid chromatography mass spectrometry (UHPLC-MS). We collected samples prior to observable disease onset and identified putative metabolite profiles that predicted future disease severity, independent of GM status. Transcriptome analyses performed after disease onset from normal epithelium and tumor tissues between the high and low tumor GMs suggests that the GM is also correlated with altered host gene expression. Integrated pathway (IP) analyses of the metabolome and transcriptome based on putatively identified metabolic features indicate that bile acid biosynthesis was enriched in rats with high tumors (GM:F344) along with increased fatty acid metabolism and mucin biosynthesis. These data emphasize the utility of using untargeted metabolomics to reveal signatures of susceptibility and resistance and integrated analysis reveals common pathways that are likely to be universal targets for intervention.
Project description:The western corn rootworm (WCR, Diabrotica virgifera virgifera LeConte) is an important pest of corn (Zea mays) in the US. Annual crop rotation between corn and soybean (Glycine max) disrupts the corn-dependent WCR lifecycle and was widely adopted to manage WCR. However, this strategy selected for a rotation-resistant (RR) variant with reduced ovipositional fidelity to cornfields. Previous studies indicated that RR-WCR adults exhibit greater tolerance of soybean tissue diet, different gut physiology, and host-microbe interactions compared to wild-types (WT). To identify genetic mechanisms underlying these phenotypic changes, a de novo assembly of the WCR adult gut transcriptome was constructed and used for RNA-sequencing analyses on RNA libraries from different WCR phenotypes (RR and WT) fed with corn or soybean diets. Differential gene expression analyses and network-based methods were used to identify gene modules transcriptionally correlated with the RR phenotype. Gene ontology enrichment analyses on these modules were then conducted to understand their potential functions and biological importance. Differential gene expression analyses on RNA libraries from adult guts of different WCR phenotypes (rotation-resistant and wild-type) fed with corn or soybean diets
Project description:Introduction : By mid-century, global atmospheric carbon dioxide concentration ([CO2]) is predicted to reach 600 umol mol-1 with global temperatures rising by 2ºC. Rising [CO2] and temperature will alter the growth and productivity of major food and forage crops across the globe. Although the impact is expected to be greatest in tropical regions, the impact of climate-change has been poorly studied in those regions. Objectives : This experiment aimed to understand the effects of elevated [CO2] (600 umol mol-1) and warming (+ 2°C), singly and in combination, on Panicum maximum Jacq. (Guinea grass) metabolite and transcript profiles. Methods: We created a de novo assembly of the Panicum maximum transcriptome. Leaf samples were taken at two time points in the Guinea grass growing season to analyze transcriptional and metabolite profiles in plants grown at ambient and elevated [CO2] and temperature, and statistical analyses were used to integrate the data. Results: The MiSeq library was quantified by qPCR and sequenced on one MiSeq flowcell for 301 cycles using paired-end sequencing. HiSeq paired-end sequencing was done with four quantified libraries per treatment which were pooled in equimolar concentration, and sequenced on two lanes for 161 cycles. The final read lengths for MiSeq and HiSeq were 300 nt and 160 nt in length. A total of 635,649,277 reads were assembled from the MiSeq/HiSeq pools. Quality control for reads generated from sequencing was performed using FastQC. Quality reads were used to perform de novo transcriptome assembly using Trinity. The initial assembly consisted of 187,216 genes. A filter was applied to keep only those genes that had at least 10 reads (across the 4 replicates) for an individual treatment. The resulting transcriptome contained 45,073 genes and reads. Functional annotation of the genes was done by using BLAST against Arabidopsis thaliana, Zea mays, and Setaria italica. Overall design: 32 sample (n=4). Panicum maximum Jacq. (Guinea Grass) was planted in a complete replicated design at University of São Paul in Ribeirão Preto, São Paulo Brazil. 4 ambient temperature and [CO2] (C) plots, 4 elevated CO2 [600 ppm] and ambient temperature (eC) plots, 4 ambient [CO2] and elevated temperature (eT) [+ 2°C] plots, and 4 combined treatment (elevated [CO2] and elevated temperature (eCeT)) plots. Each plot was 2-m in diameter; [CO2] was controlled using Free Air CO2 Enrichment and canopy warming was provided by infrared ceramic heaters as described by Britto de Assis Prado et al. (2016). Treatment was started on April 22, 2014 in a 2500 m2 field. Samples for untargeted metabolomics (with GC-MS), de novo transcriptomics, and RNASeq, were taken 30 and 50 days post treatment exposure (May 22, 2014 and July 14, 2014). Lyophilized leaf material was sent to the University of Illinois Urbana-Champagin (Ainsworth lab) for the untargeted metabolomics, transcript assembly and RNASeq.