Browse
Submit Data
Databases
API
Help

Unknown,Transcriptomics,Genomics,Proteomics

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Quantitative profiling of long noncoding RNAs with targeted RNA sequencing.

ABSTRACT: We compared the performance of conventional RNAseq with RNA Capture Sequencing (CaptureSeq) to assemble and quantify known RNA spike-Ins and human transcripts. We find CaptureSeq to be superior for the detection and quantification of the 37% lowest expressed genes, and comparable for the next 45% of moderately expressed genes. CaptureSeq contributes only minor technical variation and measures differential gene expression accurately. We demonstrate these advantages by the targeted sequencing of long noncoding RNAs across 20 human tissues, expanding previous annotations two-fold and simultaneously generating a quantitative atlas of expression. This analysis confirms the use of CaptureSeq as an important method for transcriptional profiling. Long noncoding RNA assembly and expression is analysed by targeted RNA sequencing for 20 human tissues and 4 human cell lines

ORGANISM(S): Homo sapiens

SUBMITTER: Giovanni Bussotti

PROVIDER: E-GEOD-61474 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

ACCESS DATA

Json Xml

Similar Datasets

Genome-wide discovery of human splicing branchpoints [BPCapture]

Project description:Gene splicing requires three basal genetic elements; the 3M-bM-^@M-^Y and 5M-bM-^@M-^Y splice sites and the branchpoint to which the 5M-bM-^@M-^Y intron termini is ligated to form a closed lariat during the splicing reaction. The 5M-bM-^@M-^Y and 3M-bM-^@M-^Y splice sites that define exon boundaries have been widely identified, revealing pervasive transcription and splicing of human genes. However, the locations of the third requisite element, the branchpoint, are still largely unknown. Here we employ two complementary approaches, targeted RNA sequencing and exoribonuclease digestion, to distil sequenced reads that traverse the lariat junction and, via non-conventional alignment, locate human branchpoint nucleotides. Alignments identify 88,748 branchpoints that correspond to 20% of known introns, with 76% supported by diagnostic sequence mismatch errors. This affords a first genome-wide analysis of branchpoints, describing their distribution, selection, and the existence of a diverse array of overlapping sequence motifs with distinct usage, evolutionary histories, and co-variation with distal splicing elements. The overlap of branchpoints with noncoding human genetic variation also indicates a notable contribution to disease. This annotation and analysis incorporates branchpoints into transcriptomic research and reflects a core role for this element in the regulatory code that governs gene splicing and expression. CaptureSeq identification of branchpoint nucleotides

2014-10-01 | E-GEOD-53326 | biostudies-arrayexpress

Targeted RNA sequencing for gene discovery and quantification

Project description:Due to the large size, complex splicing and wide dynamic range of eukaryotic transcriptomes, RNA sequencing samples the majority of expressed genes infrequently, resulting in sparse sequencing coverage that can hinder robust isoform assembly and quantification. Targeted RNA sequencing addresses this challenge by using oligonucleotide probes to capture selected genes or regions of interest for focused sequencing. This enhanced sequencing coverage confers sensitive gene discovery, robust transcript assembly and accurate gene quantification. Here we describe a detailed protocol for all stages of targeted RNA sequencing, from initial probe design considerations, capture of targeted genes, to final assembly and quantification of captured transcripts. Initial probe design and final analysis can take less than a day, while the central experimental capture stage requires ~7 days. Targetted RNA sequencing of long noncoding RNAs

2014-03-01 | E-GEOD-52503 | biostudies-arrayexpress

Genome-wide discovery of human splicing branchpoints

Project description:This SuperSeries is composed of the SubSeries listed below. Refer to individual Series

2014-10-01 | E-GEOD-53328 | biostudies-arrayexpress

Intergenic Disease-associated Regions are Abundant in Novel Regulatory Transcripts

Project description:Analysis of transcriptomes from 21 tissues, 13 melanoma samples and 7 breast cancer cell lines, enriched for transcripts from haploblocks with intronic and intergenic GWAS SNPs using CaptureSeq.

2017-07-04 | E-MTAB-4519 | biostudies-arrayexpress

Refining mouse annotations via targeted RNA sequencing

Project description:In this work we aim to improve the understanding of the mouse transcriptome complexity, investigate the expressed fraction of the genome and ameliorate the available mouse annotations. We utilized CatureSeq, a recently described strategy meant to enhance the sequencing coverage of low abundant genes. In our experimental design we generated oligonucleotide probes to select annotated and putative long-noncoding RNA and splice junctions. This allowed us to improve dramatically the sequencing throughput of the targeted regions. As a consequence our approach permitted the simultaneous identification of thousands of exons and the expansion of the already known ones The mouse gene assembly is anlaysed by targeted RNA sequencing of lncRNA and splice junctions. 8 mouse tissues and 16 samples are considered in the analysis. Each sample was added with external RNA controls. The controls are polyadenylated transcripts of known concentration designed to be added to an RNA analysis experiment after sample isolation.

2016-04-04 | E-GEOD-72311 | biostudies-arrayexpress

Targetted Microarray Testing for Intellectual Disability Provides Diagnoses and Identifies Novel Candidate Genes

Project description:We designed a microarray to test at exonic resolution for genomic imbalance for genes representative of all known chromosomal microdeletion/microduplication syndromes, all known causative genes for ID, all known genes encoding glutamate receptors and their known encoding proteins and all known genes encoding proteins with epigenetic regulatory function. We found 36 de novo copy number variants affecting 35 children in this study. 167 trios, each comprising an affected child and both its normal parents were analyzed by conducting two comparative hybridizations; child vs. mother and child vs. father, and only selecting a CNV if it appeared in both hybridizations, i.e., was de novo. De novo CNVs were independantly validated using quantitative PCR.

2012-07-23 | E-GEOD-39533 | biostudies-arrayexpress

Regeneration of comparative genomic hybridization oligonucleotide microarrays with dimethylurea

Project description:Reuse of materials in DNA hybridization based methods has been known since the advent of Southern membranes. Array based comparative genomic hybridization is essentially Southern hybridization with multiple probes immobilized on a solid surface. We have shown that comparative genomic hybridization microarrays fabricated with maskless array synthesizer technology can be used up to four times with application of 1,3-dimethylurea as array-stripping agent. We reproducibly detected chromosomal aberrations, 0.6 to 22.4 Mb in size, in four hybridization rounds using regenerated microarray slides. We have also demonstrated that regenerated arrays can detect smaller alterations, 16 M-bM-^@M-^S 200 kbp, such as common copy number variants, as well as complex aberration profiles in tumor. Peripheral blood leukocyte DNA samples from 7 individuals with subchromosomal aberrations vs a pool of female DNA (Promega). The experiments were performed in quadruplicate on succesively regenerated microarrays. One dye swap was performed..

2012-05-22 | E-GEOD-35740 | biostudies-arrayexpress

Expression analysis of Hypocrea jecorina CBS999.97(MAT1-2) wild-type and delta-env1 strains in four different light and darkness conditions

Project description:Investigation of whole genome gene expression level changes in response to different light conditions of the H. jecorina CBS999.97(MAT1-2) parental strain (W) and the deletion strain delta-env1 (E). These two strains were grown on malt extract agar(Merck) at 25M-bM-^DM-^C in four different conditions: (1) 24L: constant light illumination (24L); (2) 12L12D: in a 12h light/dark cycle and then 6h light illumination; (3) 12D12L: in a 12h dark/light cycle and then 6h constant darknes; (4) 24D: constant darkness. The wild-type strain is potent for sexual development in 12D12L, 12L12D and 24D, whereas the delta-env1 mutant undergo sexual development only in 24D. By contrast, the wild-type strain is female sterile in 24L, and the delta-env1 mutant is female sterile in 12D12L, 12L12D and 24D. Our results reveal that conidation-specific genes, mating locus gene, h-type maitng pheromone genes, and genes invovled in processing and secretion of h-type mating pheromone are significantly upregulated in all four female sterile conditions (W-24L, E-12L12D, E-12D12L and E-24L). We used two biological replicates of two H. jecorina CBS999.97 (MAT1-2) strains, wild-type (W) and delta-env(E) on the cellophane-covered malt extract agar (MEA) plate at 25M-bM-^DM-^C for 7.25 days.

2012-12-31 | E-GEOD-39111 | biostudies-arrayexpress

Molecular signatures of the evolving immune response in mice following a Bordetella pertussis infection.

Project description:Murine gene expression responses to Bordetella pertussis were determined in lung and spleen, between 0 and 28 days post infection. Data were compared to other parameters such as microarray, flow cytometry, multiplex immunoassays, and lung clearance. A 57 array study with 27 arrays for lung and 30 for spleen. For each tissue we used a common reference made from RNA samples of the same tissue. Please note that the reference samples are made by pooling RNA of all samples used for a tissue (either infected or uninfected). Thus, Lung and spleen each have their own reference.

2014-09-02 | E-GEOD-53294 | biostudies-arrayexpress

Effect of multiple longitudinal biopsy sampling in individual mice

Project description:Small scale study to asses the effects of taking multiple skinbiopts from individual mice in time. Results showed no influence on expression levels in time from older samplings on newer samplings. skin biopts from 4 individual mice in taken at 6 points in time

2014-09-01 | E-GEOD-51347 | biostudies-arrayexpress

OmicsDI is part of the ELIXIR infrastructure

OmicsDI is an Elixir interoperability service. Learn more ›

Tweets

OmicsDI Databases

PRIDE
PeptideAtlas
MassIVE
JPOST Repository
Physiome Model Repository

EGA
EVA
ENA
LINCS
PAXDB
Cell Collective

MetaboLights
Metabolomics Workbench
MetabolomeExpress
GNPS
BioModels
FAIRDOMHub

ArrayExpress
dbGaP
ExpressionAtlas
GEO
NODE

Information

Databases
Help
API
Contact us
Code on GitHub
Terms of use
Submit Data