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894 results for “single-cell RNA-seq”
bollito: a flexible pipeline for comprehensive single-cell RNA-seq analyses - Melanoma tutorial
<p>Downsampled version of the melanoma dataset originally published by <em><a href="https://genome.cshlp.org/content/28/9/1353">Ho et al </a>(1)</em>. The dataset is composed by cells from the 451Lu cell line. There are two samples available:</p> <table> <tbody> <tr> <td><strong>Name</strong></td> <td><strong>Description</strong></td> <td><strong>R1/R2</strong></td> </tr> <tr> <td>451LU</td> <td>Parental cell line</td> <td>2500K_451LU_L003_R*_001.fastq.gz</td> </tr> <tr> <td>451LUBR3</td> <td>Vemurafenib-resistant sample treated with targeted BRAF inhibitors</td> <td>500K_451LUBR3_L004_R*_001.fastq.gz</td> </tr> </tbody> </table> <p><br> (1) Ho YJ, Anaparthy N, Molik D, et al. Single-cell RNA-seq analysis identifies markers of resistance to targeted BRAF inhibitors in melanoma cell populations. <em>Genome Res</em>. 2018;28(9):1353-1363. doi:10.1101/gr.234062.117</p>
Single-cell RNA-seq profiles of tumor-bearing mice treated with PAGln with or without anti-PD-1
<p>single-cell RNA sequencing (scRNA-seq) profiles of tumor-bearing mice treated using Phenylacetylglutamine (PAGln) with or without anti-PD-1 were performed to compare the alterations of immune microenvironment affected by PAGln under the condition of anti-PD-1 treatment.</p>
Dataset of "Single-Cell RNA-Seq Reveals Transcriptional Heterogeneity in Latent and Reactivated HIV-infected Cells"
<p><strong>Detailed quantitative analysis of GFP expression in SAHA and TCR-treated cells & Computational analysis of bulk and single-cell RNA-Seq data.</strong></p> <p> </p> <p><em><strong>Detailed quantitative analysis of GFP expression in SAHA and TCR-treated cells.</strong></em></p> <p>Cells were prepared for single cell analysis at the Genome Technology Facility (GTF) of the University of Lausanne. Cells were loaded on Fluidigm C1 IFC plates (5-10 μm), with run ID smart33, smart34 and smart35, corresponding to untreated, SAHA- and TCR-treated conditions respectively. After single cell capture on the Fluidigm C1 IFC plate, each chamber was inspected visually by microscopy and pictures were captured with a Zeiss Axiovert 200 M fluorescence microscope equipped with a Roper Scientific CoolSnap HQ camera using a Plan-Neofluar 10X lens (smart34 run) or 20X lens (for smart35 run). For each capture chamber, pictures in bright field and FITC channel were taken with the MetaMorph 6.3 software. Picture analysis was then performed using ImageJ 1.50b software (open access software: website). Brightness and contrast were adjusted for qualitative assessment of the pictures.</p> <p><em><strong>Computational analysis of bulk and single-cell RNA-Seq data.</strong></em></p> <p>Upon bulk or single cell isolation, RNA extraction and library preparation was performed according to Illumina protocols. Bulk and single-cell RNA-Seq data analysis are detailed here.</p> <p> </p> <p>Linked to the paper published in Cell Reports (doi:10.1016/j.celrep.2018.03.102): </p> <p><strong>Single-Cell RNA-Seq Reveals Transcriptional Heterogeneity in Latent and Reactivated HIV-infected Cells</strong></p> <p>Despite effective treatment, HIV can persist in latent reservoirs, which represent a major obstacle towards HIV eradication. Targeting and reactivating latent cells is challenging due to the heterogeneous nature of HIV infected cells. Here, we used a primary model of HIV latency and single-cell RNA sequencing to characterize transcriptional heterogeneity during HIV latency and reactivation. Our analysis identified transcriptional programs leading to successful reactivation of HIV expression.</p> <p> </p> <p> </p>
Single-cell RNA-seq of breast cancer infiltrating T cells (case 1)
<p>Single cell suspensions were generated from two individual TNBC primary tumor samples (this entry contains case 2) and the viable cells were FACS sorted for CD3<sup>+</sup> T cells. Sorted cells were then counted and assessed for viability. Single cell library preparation was carried out as per the 10X Genomics Chromium Single cell protocol for the v2 reagent kit (10X Genomics, Pleasanton, CA, USA). Cell suspensions were loaded onto a Chromium Single Cell Chip along with the reverse transcription (RT) mastermix and single cell 3’ gel beads. Following generation of single cell gel bead-in-emulsions (GEMs), reverse transcription was performed using a C1000 Touch Thermal Cycler with a Deep Well Reaction Module (Bio-Rad Laboratories, Hercules, CA, USA). Amplified cDNA was purified using SPRIselect beads (Beckman Coulter, Lane Cove, NSW, Australia) and sheared to approximately 200bp with a Covaris S2 instrument (Covaris, Woburn, MA, USA) using the manufacturer’s recommended parameters. Sequencing libraries were generated with unique sample indices (SI) for each sample. Libraries were sequenced on an Illumina HiSeq 2500 High Output Mode using V4 clustering and sequencing chemistry.</p> <p>This dataset contains the raw .bcl files.</p>
Single-cell RNA-seq of breast cancer infiltrating T cells (case 2)
<p>Single cell suspensions were generated from two individual TNBC primary tumor samples (this entry contains case 1) and the viable cells were FACS sorted for CD3<sup>+</sup> T cells. Sorted cells were then counted and assessed for viability. Single cell library preparation was carried out as per the 10X Genomics Chromium Single cell protocol for the v2 reagent kit (10X Genomics, Pleasanton, CA, USA). Cell suspensions were loaded onto a Chromium Single Cell Chip along with the reverse transcription (RT) mastermix and single cell 3’ gel beads (this sample was divided into two channels). Following generation of single cell gel bead-in-emulsions (GEMs), reverse transcription was performed using a C1000 Touch Thermal Cycler with a Deep Well Reaction Module (Bio-Rad Laboratories, Hercules, CA, USA). Amplified cDNA was purified using SPRIselect beads (Beckman Coulter, Lane Cove, NSW, Australia) and sheared to approximately 200bp with a Covaris S2 instrument (Covaris, Woburn, MA, USA) using the manufacturer’s recommended parameters. Sequencing libraries were generated with unique sample indices (SI) for each sample. Libraries were sequenced on an Illumina HiSeq 2500 High Output Mode using V4 clustering and sequencing chemistry.</p> <p>This dataset contains the raw .bcl files.</p>
Mammary single-cell RNA-seq analysis and prostate cancer survival as a function of H2AFJ expression for the paper entitled: The histone variant H2A.J is enriched in luminal epithelial cells
<p>H2A.J is a poorly studied mammalian-specific variant of histone H2A. We used immunohistochemistry to study its localization in various human and mouse tissues. H2A.J showed cell-type specific expression with a striking enrichment in luminal epithelial cells of multiple glands including those of breast, prostate, pancreas, thyroid, stomach, and salivary glands. H2A.J was also highly expressed in many carcinoma cell lines and in particular, those derived from luminal breast and prostate cancer. H2A.J thus appears to be a novel marker for luminal epithelial cancers. Knocking-out the H2AFJ gene in T47D luminal breast cancer cells reduced the expression of several estrogen-responsive genes which may explain its putative tumorigenic role in luminal-B breast cancer.</p>
Joint embedding of vertebrate brain single-cell RNA-Seq using sequence or structure
<p>Embeddings of single-cell RNA-Seq data from three adult vertebrate brain datasets into Orthogroup feature space or Structural cluster feature space. Orthogroups were generated using OrthoFinder v5.5.0; Structural clusters were assigned by using FoldSeek to cluster AlphaFold-v4 structural predictions.<br> <br> The three datasets used as the basis for these embeddings were:</p> <ul> <li>sample <a href="https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSM3768152">"Brain8"</a> from the <a href="https://www.frontiersin.org/articles/10.3389/fcell.2021.743421/full">Jiang et al. 2021</a> zebrafish cell atlas (files beginning with GSM3768152)</li> <li>sample <a href="https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSM2906405">"Brain1"</a> from the <a href="https://www.sciencedirect.com/science/article/pii/S0092867418301168#sec4">Han et al. 2018</a> mouse cell atlas (files beginning with GSM2906405)</li> <li>sample <a href="https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSM6214268">"Xenopus_brain_COL65"</a> from the <a href="https://www.nature.com/articles/s41467-022-31949-2">Liao et al. 2022</a> Xenopus laevis adult cell atlas (files beginning with GSM6214268)</li> </ul> <p>For each dataset, we also generated a standardized cell type annotation file based on the author's originally provided cell type annotation data. The first column is the cell barcode for that species and the second column is the original study's cell type annotation for that cell.</p> <p>For the Xenopus brain data, we removed around ~18k cells that were not annotated in the original data to simplify data analyses - these are reflected in the files with the "subsampled" suffix. Subsampled versions of the data are also available for the joint embedding space (prefixed with "DrerMmusXlae").</p> <p>For the final datasets used in our analyses, we also provide features x cell matrices as .h5ad files for smaller file sizes and faster loading using Scanpy. </p> <p>For visualizing our UMAP plots of our top200 embedding space, we provide ".tsv" files with a variety of metrics and the x and y positions of each cell in the UMAP. See "DrerMmusXlae_adultbrain_FoldSeek_plotlydata.tsv" and "DrerMmusXlae_adultbrain_OrthoFinder_plotlydata.tsv"</p> <p>These data are part of the Arcadia Science Pub titled <a href="https://doi.org/10.57844/arcadia-vw5e-2670">"Comparing gene expression across species based on protein structure instead of sequence"</a>.</p>
Robustness and applicability of transcription factor and pathway analysis tools on single-cell RNA-seq data
<p>Data used to test the robustness and applicability of transcription factor and pathway analysis tools on single-cell RNA-seq data, described in <a href="https://doi.org/10.1186/s13059-020-1949-z">Holland et al. 2020</a>.</p> <p>The folder <em>data </em>contains<em> </em>raw data and the folder <em>output</em> contains intermediate and final results of all analyses. </p> <p>The associated analyses code and more information are available on <a href="https://github.com/saezlab/FootprintMethods_on_scRNAseq">GitHub</a>.</p> <p> </p> <p><strong>Abstract</strong></p> <p><strong>Background</strong></p> <p>Many functional analysis tools have been developed to extract functional and mechanistic insight from bulk transcriptome data. With the advent of single-cell RNA sequencing (scRNA-seq), it is in principle possible to do such an analysis for single cells. However, scRNA-seq data has characteristics such as drop-out events and low library sizes. It is thus not clear if functional TF and pathway analysis tools established for bulk sequencing can be applied to scRNA-seq in a meaningful way.</p> <p><strong>Results</strong></p> <p>To address this question, we perform benchmark studies on simulated and real scRNA-seq data. We include the bulk-RNA tools PROGENy, GO enrichment, and DoRothEA that estimate pathway and transcription factor (TF) activities, respectively, and compare them against the tools SCENIC/AUCell and metaVIPER, designed for scRNA-seq. For the in silico study, we simulate single cells from TF/pathway perturbation bulk RNA-seq experiments. We complement the simulated data with real scRNA-seq data upon CRISPR-mediated knock-out. Our benchmarks on simulated and real data reveal comparable performance to the original bulk data. Additionally, we show that the TF and pathway activities preserve cell type-specific variability by analyzing a mixture sample sequenced with 13 scRNA-seq protocols. We also provide the benchmark data for further use by the community.</p> <p><strong>Conclusions</strong></p> <p>Our analyses suggest that bulk-based functional analysis tools that use manually curated footprint gene sets can be applied to scRNA-seq data, partially outperforming dedicated single-cell tools. Furthermore, we find that the performance of functional analysis tools is more sensitive to the gene sets than to the statistic used.</p> <p> </p> <p>For questions related to the data please write an email to christian.holland@bioquant.uni-heidelberg.de or use the <a href="https://github.com/saezlab/FootprintMethods_on_scRNAseq/issues">GitHub issue system</a>.</p>
Data for "Tuning parameters of dimensionality reduction methods for single-cell RNA-seq analysis"
<p>The files named <code>df_scran.csv</code>, <code>df_seurat.csv</code>, <code>df_zinbwave.csv</code>, <code>df_dca.csv</code>, and <code>df_scvi.csv</code> contain one row per configuration that we ran successfully.</p> <p>The files named <code>DATASET.METHOD.h5ad</code> are encoded with anndata <code>v0.7.0</code> (be careful as they are not readable with previous versions) and contain 100 embeddings each. The embeddings are in the <code>obsm</code> attribute of the object. All the embeddings can be listed with the <code>obsm_keys()</code> method. The name of the embedding contains the parameters used to generate that embedding and are written like that <code>method=zinbwave.dims=10.epsilon=1000.features=300.gene_covariate=0</code>.</p> <p> </p> <p>For questions on this dataset please contact fraimundo@google.com</p>
EI Single-Cell RNA-Seq Workshop 2020
<p>Datasets to be used for the "Single-Cell RNA-Seq Workshop 2020" at the Earlham Institute, Norwich, UK.</p>
Beyondcell: targeting cancer therapeutic heterogeneity in single-cell RNA-seq
<p><strong><a href="https://gitlab.com/bu_cnio/Beyondcell">Beyondcell</a> </strong>is a methodology for the identification of drug vulnerabilities in single cell RNA-seq data. To this end, <strong>Beyondcell</strong> focuses on the analysis of drug-related commonalities between cells by classifying them into distinct therapeutic clusters. We have validated the tool in a population of MCF7-AA cells exposed to 500nM of bortezomib and collected at different time points: t0 (before treatment), t12, t48 and t96 (72h treatment followed by drug wash and 24h of recovery) obtained from <a href="https://www.nature.com/articles/s41586-018-0409-3"><strong><em>Ben-David U, et al., Nature, 2018</em></strong></a>. Here, you can find the integrated Seurat object obtained from this analysis. This object is meant to help users follow <strong>Beyondcell's</strong> <a href="https://gitlab.com/bu_cnio/Beyondcell/-/tree/master/tutorial/analysis_workflow">analysis workflow</a>.</p> <p> </p>
Single-cell RNA-seq profiles of lung adenocarcinoma patients and tumor-bearing mice
<p>single-cell RNA sequencing (scRNA-seq) profiles from eight patients with lung adenocarcinoma (LUAD) and four samples of tumor tissues from tumor bearing mice were performed. By integrating other scRNA-seq data and clinical information, we identified activated adaptive immune responses in older patients, reflected by enriched dysfunctional T cell signature scores and immune checkpoint molecules. Our study shows increased efficacy of immune checkpoint blockade therapy in older patients, addressing the prominent role of age when considering immunotherapy.</p>
A Galaxy-based training resource for single-cell RNA-seq quality control and analyses
<p>This is the tutorial data for the 'Single-cell quality control with scater' tutorial on the Galaxy Training Network. The data is the same dataset that is used as the inbuilt example dataset within scater, but has been implemented as individual files.</p>
Processed data for "Dissociation of solid tumour tissues with cold active protease for single-cell RNA-seq minimizes conserved collagenase-associated stress responses"
<p>tar.gz of processed data in the form of compressed R files (rds) of SingleCellExperiment (<a href="https://bioconductor.org/packages/release/bioc/html/SingleCellExperiment.html">https://bioconductor.org/packages/release/bioc/html/SingleCellExperiment.html</a>) objects and a metadata csv for the data in the publication <em>Dissociation of solid tumour tissues with cold active protease for single-cell RNA-seq minimizes conserved collagenase-associated stress responses </em>(O'Flanagan et al. 2019).</p>
Supplementary Data: Identification of experimentally-supported poly(A) sites in single-cell RNA-seq data with SCINPAS
<p>Supplementary data for the SCINPAS analysis as part of the manuscript "Identification of experimentally-supported poly(A) sites in single-cell RNA-seq data with SCINPAS".</p> <p>It includes the output of SCINPAS, available here: <a href="https://github.com/zavolanlab/SCINPAS">https://github.com/zavolanlab/SCINPAS</a>, used for the analyses. Additionally, the scripts and data for the comparison with other tools is provided. Please read the README.md for more information.</p>
Generating an expression matrix for droplet single-cell RNA-seq (dscRNA-seq) data
<p>This tutorial is adapted from the 'Generating an expression matrix' training session at the EBI (https://www.ebi.ac.uk/training/events/2019/single-cell-rna-seq-analysis-questions-clusters).</p>
Datasets associated with the manuscript "Differential detection workflows for multi-sample single-cell RNA-seq data"
<p>In this Zenodo repository, we share the data that is required to reproduce all the analyses from our publication "Differential detection workflows for multi-sample single-cell RNA-seq data".</p> <p>This repository includes all* input data, intermediate results and final outputs that are represented in our manuscript. For a more elaborate description of the data, we refer to the companion GitHub. https://github.com/statOmics/DD_benchmarks for the benchmarks and https://github.com/statOmics/DD_cases for the case studies, respectively.</p>
Additional data for manuscript "Alevin-fry unlocks rapid, accurate, and memory-frugal quantification of single-cell RNA-seq data"
<p>Additional data for manuscript "Alevin-fry unlocks rapid, accurate, and memory-frugal quantification of single-cell RNA-seq data".</p> <p>Additional mitochondrial gene sequences for Danio rerio, Homo sapiens, and Mus musculus.</p>
Processing single-cell RNA-seq datasets using SingCellaR
<ul> <li>cellranger_output.zip : Zipped files for cellranger output</li> <li>Code.zip: This contains the code for Step2-7</li> <li>Human_genesets.zip: This includes the geneset signatures (.gmt) files we used in the protocol and original study (Roy et al, 2021)</li> <li>Human_HSPC_All.SingCellaR.rdata: The SingCellaR objects generated in Step 4 on a local computer </li> <li>ABM_1.SingCellaR.rdata: The SingCellaR objects generated in Step 3 on a local computer </li> <li>eFL_All.SingCellaR.rdata: The SingCellaR objects generated in Step 3 on a local computer </li> <li>FBM_All.SingCellaR.rdata: The SingCellaR objects generated in Step 3 on a local computer </li> <li>FL_All.SingCellaR.rdata: The SingCellaR objects generated in Step 3 on a local computer </li> <li>PBM_All.SingCellaR.rdata: The SingCellaR objects generated in Step 3 on a local computer </li> <li>meta.data.txt: meta data includes the donor and batch information </li> </ul>
Identifying cell states in single-cell RNA-seq data at statistically maximal resolution
<p>In this repository we provide the datasets for the results of the Cellstates method as shown in the article: “Identifying cell states in single-cell RNA-seq data at statistically maximal resolution” by Pascal Grobecker, Thomas Sakoparnig, and Erik van Nimwegen.</p> <p>A preprint is available under the following DOI: https://doi.org/10.1101/2023.10.31.564980</p> <p>There is a README file that describes the contents and formats of each of the tab-separated values files. There is one subdirectory containing files with Cellstates' results on the dataset of Zeisel et al. (DOI: 10.1016/j.cell.2018.06.021) which is accompanied by another README file describing the formats and contents of these files. </p>
ScienceDex guides
Understand access before you commit
These curated guides explain access requirements, typical timelines, costs, and reuse considerations for widely used research datasets.
Allen Brain Atlas
Allen Brain Atlas is an Allen Institute collection of brain map atlases, datasets, APIs, and analysis tools covering mouse, human, and non-human primate brain resources.
Annotated Behaviour and Observability Dataset (ABODe)
ABODe is a University of Edinburgh DataShare dataset for behavior classification in group-housed mice using home-cage video, identities, bounding boxes, ground-plate positions, and annotator labels.
DANDI Archive for NWB datasets
DANDI is a BRAIN Initiative archive for publishing and sharing neurophysiology data, including electrophysiology, optophysiology, and behavioral data packaged as NWB and related standards.
International Brain Laboratory public data
The International Brain Laboratory public data releases expose standardized mouse decision-making experiments, including Neuropixels recordings, widefield calcium imaging, behavior, and session metadata accessed through the ONE API.
OpenNeuro
OpenNeuro is a free, open platform for sharing neuroimaging datasets, with public search, dataset pages, and download paths for web, S3, DataLad, and the OpenNeuro CLI.