Project description:we developed a low-input and user-friendly pipeline for the simultaneous profiling of five distinct Cys states with regional resolution in the gut of monkeys, including free SH, total Cys oxidation (Sto), sulfenic acid (SOH), S-nitrosylation (SNO), and S-glutathionylation (SSG), shedding light on redox modification stoichiometries and redox signaling of gut aging.Furthermore, redox-modification and proteomic analysis of colonic tissues of calorie-restricted aged mice revealed that CR can reduce oxidative stress and highlight the protective effect of CR on intestinal health.
Project description:The increasing application of RNA-seq to study non-model species demands easy-to-use and efficient bioinformatics tools to help researchers quickly uncover biological and functional insights. We developed ExpressAnalyst (www.expressanalyst.ca), a web-based tool for processing, analyzing, and interpreting RNA-seq data from any eukaryotic species. ExpressAnalyst contains a series of modules that enable raw data processing and annotation of FASTQ files, and statistical and functional analysis of counts tables and gene lists. All modules are integrated with EcoOmicsDB, an ortholog database that enables comprehensive analysis for species without a reference transcriptome. By coupling ultra-fast read mapping algorithms with high-resolution ortholog databases through a user-friendly web interface, ExpressAnalyst enables researchers to obtain global expression profiles and gene-level insights from raw RNA-seq reads within 24 hours. Here, we present ExpressAnalyst and demonstrate its utility with a case study of RNA-seq data from multiple non-model salamander species, including two that do not have a reference transcriptome.
Project description:The present work describes a novel search program for the detection of both MS-cleavable and non-cleavable crosslinked peptides and it is the first software program reported with both capacities. The search strategy has been implemented in the computer program MetaMorpheus, which has a user-friendly graphical user interface (GUI). Novel crosslinker molecules can be easily added if desired. A fragment-ion index scheme makes the search computationally efficient. DSSO crosslinked BSA and E.coli ribosome data were used to validate the MS-cleavable crosslink search results.
2018-07-16 | PXD009000 | Pride
Project description:Development of a user-friendly pipeline for mutational anal-yses of HIV using ultra-accurate maximum-depth sequencing
Project description:Here, we present ChromSCape, a user-friendly interactive Shiny/R application that processes single-cell epigenomic data to help the biological interpretation of chromatin landscapes within cell populations. ChromSCape successfully analyses the distribution of repressive and active histone modifications as well as chromatin accessibility landscapes from single-cell datasets.
Project description:We present GuideScan2 for memory-efficient, parallelizable construction of high-specificity CRISPR guide RNA (gRNA) databases and user-friendly design and analysis of individual gRNAs and gRNA libraries for targeting coding and non-coding regions in custom genomes. GuideScan2 analysis identifies widespread confounding effects of low-specificity gRNAs in published CRISPR screens and enables construction of a gRNA library that reduces off-target effects in a gene essentiality screen. GuideScan2 also enables the design and experimental validation of allele-specific gRNAs in a hybrid mouse genome. GuideScan2 will facilitate CRISPR experiments across a wide range of applications.
Project description:Circadian clocks are important for gut health. This experiment aimed to determine the role of core clock gene Bmal1 in regulating microbial rhythmicity in health and dextran sulphate sodium induced colitis. Mice were generated with Bmal1 selectively deleted in Villin-expressing cells (predominantly IECs).Microbial DNA was extracted from fecal pellets collected from IEC-Bmal1-/- and Bmal1flox mice (aged 8-19 weeks) at zeitgeber time 0, 4, 8, 12, 16, 20 across the 24h day with the DNeasy PowerSoil Pro Kit (Qiagen), as per manufacturer’s instructions. Pre-amplification of the V4 region of 16S rRNA was performed using forward primer 5'-ACACTCTTTCCCTACACGACGCTCTTCCGAT-CTNNNNNGTGCCAGCMGCCGCGGTAA-3' (annealing sites in bold) and reverse primer 5'-GTGACTGGAGTTCAGACGTGTGCTCTTCCGATCTGGACTACHVGGGTWTCTAAT-3'. Sequencing was performed by the University of Liverpool Centre for genomics Research, using the Illumina MiSeq v2 platform (Illumina), generating 250bp paired-end reads. PhiX control v3 library (PhiX) was spiked into samples to balance low base diversity often found in microbiome samples. Quality control was performed and OTU tables were generated using a pipeline provided by the University of Manchester Bioinformatics Core Facility local Galaxy service. Briefly, VSEARCH clustered OTUs and removed chimeras. The OTU database was mapped to the SILVA (v138) reference database with >97% homology threshold. All samples passed quality checks and had sequence depth >45,000. The OTU table was analysed using R packages phyloseq, vegan, limma and ALDEx2. JTK_CYCLE 93 was used to identify rhythmic OTUs with a period of 24 h and an adjusted P value < 0.05.
Project description:Background Single-cell RNA-sequencing (scRNA-seq) experiments typically analyze hundreds or thousands of cells after amplification of the cDNA. The high throughput is made possible by the early introduction of sample-specific bar codes (BCs), and the amplification bias is alleviated by unique molecular identifiers (UMIs). Thus, the ideal analysis pipeline for scRNA-seq data needs to efficiently tabulate reads according to both BC and UMI. Findings zUMIs is a pipeline that can handle both known and random BCs and also efficiently collapse UMIs, either just for exon mapping reads or for both exon and intron mapping reads. If BC annotation is missing, zUMIs can accurately detect intact cells from the distribution of sequencing reads. Another unique feature of zUMIs is the adaptive downsampling function that facilitates dealing with hugely varying library sizes but also allows the user to evaluate whether the library has been sequenced to saturation. To illustrate the utility of zUMIs, we analyzed a single-nucleus RNA-seq dataset and show that more than 35% of all reads map to introns. Also, we show that these intronic reads are informative about expression levels, significantly increasing the number of detected genes and improving the cluster resolution. Conclusions zUMIs flexibility makes if possible to accommodate data generated with any of the major scRNA-seq protocols that use BCs and UMIs and is the most feature-rich, fast, and user-friendly pipeline to process such scRNA-seq data.