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ShareScore release 0.9.0
Dataset results
36 results for “single molecule RNA”
Single-molecule Fluorescent In Situ Hybridization (smFISH) for RNA detection in the fungal pathogen Candida albicans dataset
<p><strong>This dataset is connected to the protocol article titled:</strong></p> <p>Single-molecule Fluorescent <em>In Situ</em> Hybridization (smFISH) for RNA detection in the fungal pathogen <em>Candida albicans</em></p> <p><strong>Abstract:</strong></p> <p><em>Candida albicans</em> is the most prevalent human fungal pathogen. Its pathogenicity is linked to the ability of <em>C. albicans</em> to reversibly change morphology and to grow as yeast, pseudohyphal or hyphal cells in response to environmental stimuli. Understanding the molecular regulation controlling those morphological switches remains a challenge that, if solved, could help fight <em>C. albicans</em> infections.</p> <p>While numerous studies investigated gene expression changes occurring during <em>C. albicans</em> morphological switches using bulk approaches (e.g., RNA sequencing), here we describe a single-cell and single-molecule RNA imaging and analysis protocol to measure absolute mRNA counts in morphologically intact cells. To detect endogenous mRNAs in single fixed cells, we optimized a single molecule fluorescent <em>in situ</em> hybridization (smFISH) protocol for <em>C. albicans</em>, which allows one to quantify the differential expression of mRNAs in yeast, pseudohyphae or hyphal cells. We quantified the expression of two mRNAs, cell cycle-controlled mRNA (<em>CLB2)</em> and a transcription regulator (<em>EFG1</em>), which show differential expression in the different morphological cell types and in different nutrient conditions. In this protocol we described in detail the major steps of this approach: growth and fixation, hybridization, imaging, cell-segmentation and mRNA spot analysis. Raw data is provided with the protocol to favour reproducibility. This approach could benefit the molecular characterization of <em>C. albicans</em> and other filamentous fungi, pathogenic or non-pathogenic.</p> <p><strong>Data description:</strong></p> <p>This dataset consists of a FISH experiment spanning two different mRNAs, EFG1 and CLB2, and two different nutrient condition, being SPIDER37 and TSB37 in Candida albicans. For culturing, the C. albicans wildtype strain SC5314 was inoculated at 30 degrees overnight (~15 hours) in 10 mL of TSB medium in a 30 degree (celsius) shaking incubator. Next, samples were diluted to a density of 10^5 cells/ mL and inoculated for 6 hours in either 30 mL TSB medium or Spider medium at 37 degrees in falcon tubes on an orbital microplate shaker. Then, samples were fixated by adding PFA to a final concentration of 4% to the medium. For hybridization, both mRNAs were hybridized independently by specific DNA oligo labelled with a Quasar670 dye to enable the visualisation of single mRNA molecules. As both genes are labelled by the same dye, these oligos were not co-applied to the same sample but to independent samples.</p> <p><strong>Microscopy</strong></p> <p>For smFISH imaging we use an Olympus BX-63 epifluorescence microscope equipped with Ultrasonic stage and UPlanApo 100x 1.35NA oil-immersion objective (Olympus). Lumencore SOLA FISH light source, a Hamamatsu ORCA-Fusion sCMOS camera (6.5 µm-pixel size) mounted using U-CMT C-Mount Adapter, and zero-pixel shift filter sets: F36-500 DAPI HC Brightline Bandpass Filter, F36-502 FITC HC BrightLine Filter, F36-542 Cy3 HC BrightLine Filter, and F36-523 Cy5 HC BrightLine Filter. Images are acquired across 61-81 optical sections (depending on the sample thickness) with a z-step size of 0.2 μm. The CellSens software (Olympus) is used for instrument control and image acquisition. For the DAPI channel 10-50 ms of exposure was used. Whilst, for the CY5 channel, used imaging the FISH probes, 750 ms was applied. </p> <p> </p>
Single-molecule Fluorescent In Situ Hybridization (smFISH) for RNA detection in the fungal pathogen Candida albicans small example dataset
<p><strong>This small example dataset is connected to the protocol article titled:</strong></p> <p>Single-molecule Fluorescent <em>In Situ</em> Hybridization (smFISH) for RNA detection in the fungal pathogen <em>Candida albicans</em></p> <p><strong>Abstract:</strong></p> <p><em>Candida albicans</em> is the most prevalent human fungal pathogen. Its pathogenicity is linked to the ability of <em>C. albicans</em> to reversibly change morphology and to grow as yeast, pseudohyphal or hyphal cells in response to environmental stimuli. Understanding the molecular regulation controlling those morphological switches remains a challenge that, if solved, could help fight <em>C. albicans</em> infections.</p> <p>While numerous studies investigated gene expression changes occurring during <em>C. albicans</em> morphological switches using bulk approaches (e.g., RNA sequencing), here we describe a single-cell and single-molecule RNA imaging and analysis protocol to measure absolute mRNA counts in morphologically intact cells. To detect endogenous mRNAs in single fixed cells, we optimized a single molecule fluorescent <em>in situ</em> hybridization (smFISH) protocol for <em>C. albicans</em>, which allows one to quantify the differential expression of mRNAs in yeast, pseudohyphae or hyphal cells. We quantified the expression of two mRNAs, cell cycle-controlled mRNA (<em>CLB2)</em> and a transcription regulator (<em>EFG1</em>), which show differential expression in the different morphological cell types and in different nutrient conditions. In this protocol, we described in detail the major steps of this approach: growth and fixation, hybridization, imaging, cell-segmentation and mRNA spot analysis. Raw data is provided with the protocol to favour reproducibility. This approach could benefit the molecular characterization of <em>C. albicans</em> and other filamentous fungi, pathogenic or non-pathogenic.</p> <p><strong>Data description:</strong></p> <p>This dataset consists of a FISH experiment spanning two different mRNAs, EFG1 and CLB2, and one nutrient condition, SPIDER37, in Candida albicans. For culturing, the C. albicans wildtype strain SC5314 was inoculated at 30 degrees overnight (~15 hours) in 10 mL of TSB medium in a 30 °C shaking incubator. Next, samples were diluted to a density of 10^5 cells/ mL and inoculated for 6 hours in 30 mL Spider medium at 37 °C in falcon tubes on an orbital microplate shaker. Then, samples were fixated by adding PFA to a final concentration of 4% to the medium. For hybridization, both mRNAs were hybridized independently by specific DNA oligo labelled with a Quasar670 dye to enable the visualisation of single mRNA molecules. As both genes are labelled by the same dye, these oligos were not co-applied to the same sample but to independent samples.</p> <p><strong>Microscopy</strong></p> <p>For smFISH imaging we use an Olympus BX-63 epifluorescence microscope equipped with Ultrasonic stage and UPlanApo 100x 1.35NA oil-immersion objective (Olympus). Lumencore SOLA FISH light source, a Hamamatsu ORCA-Fusion sCMOS camera (6.5 µm-pixel size) mounted using U-CMT C-Mount Adapter, and zero-pixel shift filter sets: F36-500 DAPI HC Brightline Bandpass Filter, F36-502 FITC HC BrightLine Filter, F36-542 Cy3 HC BrightLine Filter, and F36-523 Cy5 HC BrightLine Filter. Images are acquired across 61-81 optical sections (depending on the sample thickness) with a z-step size of 0.2 μm. The CellSens software (Olympus) is used for instrument control and image acquisition. For the DAPI channel 10-50 ms of exposure was used. Whilst, for the CY5 channel, used for imaging the FISH probes, 750 ms was applied. </p>
Reference genes for quantitative Arabidopsis single molecule RNA fluorescence in situ hybridization
Abstract Subcellular mRNA quantities and spatial distributions are fundamental for driving gene regulatory programmes. Single molecule RNA fluorescence in situ hybridization (smFISH) uses fluorescent probes to label individual mRNA molecules, thereby facilitating both localization and quantitative studies. Validated reference mRNAs function as positive controls and are required for calibration. Here we present selection criteria for the first set of Arabidopsis smFISH reference genes. Following sequence and transcript data assessments, four mRNA probe sets were selected for imaging. Transcript counts per cell, correlations with cell size, and corrected fluorescence intensities were all calculated for comparison. In addition to validating reference probe sets, we present sample preparation steps that can retain green fluorescent protein fluorescence, thereby providing a method for simultaneous RNA and protein detection. In summary, our reference gene analyses, modified protocol, and simplified quantification method together provide a firm foundation for future quantitative single molecule RNA studies in Arabidopsis root apical meristem cells.
Reference genes for quantitative Arabidopsis single molecule RNA fluorescence in situ hybridization
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single-molecule tertiary structure determination reveals RNA folding landscape
<p>Initial release of the 6HB RNA origami 3D density maps, fitting models and EMDB validation reports.</p>
Data from: Single-molecule RNA detection at depth by hybridization chain reaction and tissue hydrogel embedding and clearing
Open the record for dataset details and reuse information.
Comparative analysis between single-cell RNA-seq and single-molecule RNA FISH indicates that the pyrimidine nucleobase idoxuridine (IdU) globally amplifies transcriptional noise
GEO Series GSE263194. Homo sapiens. 4 samples. Type: Expression profiling by high throughput sequencing.
A comparison between single cell RNA sequencing and single molecule RNA FISH for rare cell analysis
GEO Series GSE99330. Homo sapiens. 801 samples. Type: Expression profiling by high throughput sequencing.
Directed protein evolution of an RNA editor for single molecule RNA-binding protein co-occupancy (I)
GEO Series GSE216334. Homo sapiens. 138 samples. Type: Other.
Digital RNA Sequencing Minimizes Sequence-Dependent Bias and Amplification Noise with Optimized Single Molecule Barcodes
GEO Series GSE34449. Escherichia coli str. K-12 substr. MG1655. 2 samples. Type: Expression profiling by high throughput sequencing.
Single-molecule identification of the target RNAs of different RNA binding proteins simultaneously in cells
GEO Series GSE200711. Homo sapiens. 54 samples. Type: Other.
Genome-wide single molecule footprinting reveals high RNA polymerase II turnover at paused promoters
GEO Series GSE77369. Drosophila melanogaster. 76 samples. Type: Methylation profiling by high throughput sequencing; Non-coding RNA profiling by high throughput sequencing.
Directed protein evolution of an RNA editor for single molecule RNA-binding protein co-occupancy (II)
GEO Series GSE241615. Homo sapiens. 23 samples. Type: Other.
Single-cell RNA-seq for identification of B-cell specific checkpoint molecules regulating anti-tumor immunity
GEO Series GSE225713. Mus musculus. 40 samples. Type: Expression profiling by high throughput sequencing.
High-throughput single-molecule long-read RNA sequencing analysis of tissue-specific genes and isoforms in lettuce (Lactuca sativa L.)
GEO Series GSE264560. Lactuca sativa. 12 samples. Type: Expression profiling by high throughput sequencing.
Single-molecule chromatin configurations link transcription factor binding to expression in human cells [RNA-seq]
GEO Series GSE276515. Homo sapiens. 4 samples. Type: Expression profiling by high throughput sequencing.
Single-molecule RNA-FISH analysis reveals stochasticity in reactivation of latent HIV-1 regulated by Nuclear Orphan Receptors NR4A and cMYC
GEO Series GSE241207. Homo sapiens. 3 samples. Type: Expression profiling by high throughput sequencing.
Concerted modification of nucleotides at functional centers of the ribosome revealed by single-molecule RNA modification profiling
GEO Series GSE186634. Saccharomyces cerevisiae. 33 samples. Type: Expression profiling by high throughput sequencing.
Single-Molecule Direct RNA Sequencing Reveals the Shaping of Epitranscriptome Across Multiple Species
GEO Series GSE246632. Chlamydomonas reinhardtii; Arabidopsis thaliana; Mus musculus; Oryza sativa; Homo sapiens. 13 samples. Type: Expression profiling by high throughput sequencing; Methylation profiling by high throughput sequencing.
Single cell RNA sequencing of human induced pluripotent stem cell-derived cardiomyocytes with or without small molecule activation of FOXO signaling
GEO Series GSE190742. Homo sapiens. 4 samples. Type: Expression profiling by high throughput sequencing.
ScienceDex guides
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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.