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

Bulk RNA Barcoding and sequencing (BRB-seq) protocol for RNA-seq of 60 human LCL from the 1000G project

ABSTRACT: Here, we validate a novel protocol, Bulk RNA Barcoding and sequencing (BRB-seq), that combines the multiplexing-driven cost-effectiveness of a single-cell RNA-seq protocol with the efficiency of a bulk RNA-seq procedure. For this we use BRB-seq protocol on 60 human LCL data from the 1000G project samples, in order to show its multiplexing capacity and compare its effectiveness to already published TruSeq results on same samples.

INSTRUMENT(S): NextSeq 500

ORGANISM(S): Homo sapiens

SUBMITTER: Vincent Gardeux

PROVIDER: E-MTAB-7524 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

ACCESS DATA

Json Xml

Publications

BRB-seq: ultra-affordable high-throughput transcriptomics enabled by bulk RNA barcoding and sequencing.

Alpern Daniel D Gardeux Vincent V Russeil Julie J Mangeat Bastien B Meireles-Filho Antonio C A ACA Breysse Romane R Hacker David D Deplancke Bart B

Genome biology 20190419 1

Despite its widespread use, RNA-seq is still too laborious and expensive to replace RT-qPCR as the default gene expression analysis method. We present a novel approach, BRB-seq, which uses early multiplexing to produce 3' cDNA libraries for dozens of samples, requiring just 2 hours of hands-on time. BRB-seq has a comparable performance to the standard TruSeq approach while showing greater tolerance for lower RNA quality and being up to 25 times cheaper. We anticipate that BRB-seq will transform ...[more]

PMID: 30999927

Similar Datasets

RNA-sequencing of 465 lymphoblastoid cell lines from the 1000 Genomes

Project description:10.1038/nature12531

2012-11-06 | E-GEUV-1 | biostudies-arrayexpress

Re-analysis of 667 assays from the GEUVADIS study

Project description:Re-analysis of 667 assays (463 samples) from Geuavdis study has been performed to demonstrate the flexibility of the Ballgown package to identify transcript-level eQTLs and identify non-linear artifacts in transcript data. Our package Ballgown is freely available from: https://github.com/alyssafrazee/ballgown. Geuvadis RNA sequencing data set of 465 human lymphoblastoid cell line samples from the CEU, FIN, GBR, TSI and YRI populations from the 1000 Genomes sample collection was created by the Geuvadis consortium (www.geuvadis.org, http://www.geuvadis.org/web/geuvadis/our-rnaseq-project). Original Geuvadis mRNA and small RNA sequencing data, clean data that passed QC and other filters, processed files and analysis results are available under accession E-GEUV-1, E-GEUV-2, E-GEUV-3.

2014-04-17 | E-GEUV-6 | biostudies-arrayexpress

small RNA-sequencing of 452 lymphoblastoid cell lines from the 1000 Genomes

Project description:This RNA sequencing data set of 465 human lymphoblastoid cell line samples from the CEU, FIN, GBR, TSI and YRI populations from the 1000 Genomes sample collection was created by the Geuvadis consortium (www.geuvadis.org, http://www.geuvadis.org/web/geuvadis/our-rnaseq-project). The data is under embargo until the first publication by the investigators in early 2013. This accession contains small RNA sequencing data, and mRNA data from the same samples are available under accession E-GEUV-1.

2012-11-06 | E-GEUV-2 | biostudies-arrayexpress

RNA-sequencing and small RNA-sequencing of 465 lymphoblastoid cell lines from the 1000 Genomes

Project description:This RNA sequencing data set of 465 human lymphoblastoid cell line samples from the CEU, FIN, GBR, TSI and YRI populations from the 1000 Genomes sample collection was created by the Geuvadis consortium (www.geuvadis.org, http://www.geuvadis.org/web/geuvadis/our-rnaseq-project). The data is under embargo until the first publication by the investigators in early 2013. This accession contains all mRNA and small RNA sequencing data, clean data that passed QC and other filters are available under accession E-GEUV-1, E-GEUV-2.

2012-12-06 | E-GEUV-3 | biostudies-arrayexpress

Impact of Regulatory Variation from RNA to Protein

Project description:Genetic variants that impact gene regulation are important contributors to human phenotypic variation. For this reason, considerable efforts have been made to identify genetic associations with differences in mRNA levels of nearby genes, namely, cis expression quantitative trait loci (eQTLs). The phenotypic consequences of eQTLs are presumably due, in most cases, to their ultimate effects on protein expression levels. Yet, only few studies have quantified the impact of genetic variation on proteins levels directly. It remains unclear how faithfully eQTLs are reflected at the protein level, and whether there is a significant layer of cis regulatory variation acting primarily on translation or steady state protein levels. To address these questions, we measured ribosome occupancy by high-throughput sequencing, and relative protein levels by high-resolution quantitative mass spectrometry, in a panel of lymphoblastoid cell lines (LCLs) in which we had previously measured transcript expression using RNA sequencing. We then mapped genetic variants that are associated with changes in transcript expression (eQTLs), ribosome occupancy (rQTLs), or protein abundance (pQTLs). Most of the QTLs we detected are associated with transcript expression levels, with consequent effects on ribosome and protein levels. However, we found that eQTLs tend to have significantly reduced effect sizes on protein levels, suggesting that their potential impact on downstream phenotypes is often attenuated or buffered. Additionally, we confirmed the presence of a class of cis QTLs that specifically affect protein abundance with little or no effect on mRNA levels; most of these QTLs have little effect on ribosome occupancy, and hence may arise from differences in post-translational regulation.

2014-12-19 | PXD001406 | Pride

A Methodological Assessment and Characterization of Genetically-Driven Variation in Three Human Phosphoproteomes

Project description:Phosphorylation of proteins on serine, threonine, and tyrosine residues is a ubiquitous post-translational modification that plays a key part of essentially every cell signaling process. It is reasonable to assume that inter-individual variation in protein phosphorylation may underlie phenotypic differences, as has been observed for practically any other molecular regulatory phenotype. However, we do not know much about the extent of inter-individual variation in phosphorylation because it is quite challenging to perform a quantitative high throughput study to assess inter-individual variation in any post-translational modification. To test our ability to address this challenge with current technology, we quantified phosphorylation levels for three fully sequenced human cell lines within a nested experimental framework, and found that genetic background is the primary determinant of phosphoproteome variation. We uncovered multiple functional, biophysical, and genetic associations with germline driven phosphopeptide variation (though the small sample size in this ‘pilot’ study limits the applicability of our genetic observations). Among these associations were variants affecting protein levels or structure, with the latter presenting, on average, a stronger effect. Interestingly, we found evidence that is consistent with a phosphopeptide variability buffering effect endowed from properties enriched within longer proteins. We also undertook a thorough technical assessment of our experimental workflow to aid further efforts. Taken together, this work provides the foundation for future work to characterize inter-individual variation in post-translational modification as well as reveals novel insights into the nature of inter-individual variation in phosphorylation.

2018-10-22 | PXD008002 | Pride

Glucocorticoid treatment leads to aberrant ion and macromolecular transport mechanisms in regenerating zebrafish fins.

Project description:Bone is a highly regenerative tissue but it can be permanently lost due to severe trauma and as a result of disease. In zebrafish, which are capable of complex organ regeneration, intramembranous bones of the fins and skull regenerate rapidly even after profound loss. The fin regenerate, albeit being composed of different tissues is a structure of modest complexity which, together with the fact that it is easy to access and monitor, makes it a useful system to study bone formation, wound healing and metabolic disease. Glucocorticoids are powerful drugs used in anti-inflammatory and immunosuppressive therapy. Despite their unmistakable advantages and usefulness, long term treatment with glucocorticoids can cause a variety of adverse effects such as insulin resistance, high blood pressure and bone fragility. Here we made use of proteomics to study the fin regeneration by sampling zebrafish caudal fins without prior injury and regenerated tissue 4 days post amputation at normal and perturbed conditions (prednisolone-treatment).

2019-09-26 | PXD013552 | Pride

RNA-seq of proximal, middle, and distal caudal fin tissue of thyroid-ablated zebrafish (hypothyroid, HypoTH) and wild-type zebrafish (euthyroid, EuTH)

Project description:The goal of this experiment is to test the hypothesis that hypothyroid zebrafish caudal fin possess proximalized gene expression profile. Transcriptome data was generated for proximal, middle and distal adult fin tissue of hypothyroid zebrafish, with euthyroid sibling fish as control.

2022-04-12 | E-MTAB-9528 | biostudies-arrayexpress

Transcription profiling of Zebrafish 1 Day post amputation vehicle or beclomethasone exposed to study regeneration

Project description:Zebrafish caudal fin regeneration is an established model to study tissue regeneration. In order to identify novel molecular signaling pathways critical for regeneration, we developed a rapid throughput in vivo regeneration assay. We screened a 2000 member structurally diverse small molecule library, followed by assessment of regenerative progression at three days post amputation. A cluster of glucocorticoids was identified among the âpositive hitsâ. To identify the molecular targets of the activated glucocorticoid receptor, microarray analysis was performed using RNA isolated from the regenerates of control and glucocorticoid exposed zebrafish. We identified 673 transcripts that were differentially regulated. The level of expression and spatial expression pattern of select genes were completed by qPCR and by in situ hybridization, respectively. Altogether, these studies demonstrate the power of chemical genetics to identify chemical probes and their targets which will provide a path towards defining conserved regenerative pathways. Experiment Overall Design: The caudal fin of zebrafish larvae at 2days post fertilization were amputated and exposed to vehicle control alone or Beclomethasone . Regenerating fins were isolated at 1days post amputation. Three replicates were collected at each time point. 150 fins were pooled to comprise one replicate.

2009-02-08 | E-GEOD-10766 | biostudies-arrayexpress

Functional effect of DOT1L pharmacological inhibition in ERα-positive Ovarian Cancer cells

Project description:Estrogen Receptor alpha (ERα), a nuclear receptor with transcriptional activity, is a master regulator of estrogen signaling, widely known as therapeutic target in hormone-responsive breast cancer (BC). Moreover, ERα is highly expressed in approximately 80% of High Grade Serous Ovarian Cancer (HGSOC), the most common epithelial ovarian carcinoma. Despite some promising clinical trials evaluating endocrine therapy in this type of tumor, the role of ERα is still unknown. Epigenetic changes, such as DNA methylation, are emerging as key contributing factors to carcinogenesis. Disruptor of telomeric silencing-1-like (DOT1L), the only known histone methyl transferase capable to produce H3K79 mono, di and tri-methylation, modulates ERα actions in hormone-responsive BC. Considering this evidence, ERα-DOT1L association was confirmed in ERα-positive OC cells, PEO1 and PEO4, by Co-IP. DOT1L pharmacological inhibition by EPZ004777 (EPZ) revealed the involvement of this epigenetic enzyme in cell proliferation, cell cycle progression and apoptosis. Transcriptome profiling after ICI (a Selective Estrogen Receptor Degrader) and EPZ treatment, in both cell lines, has underlined a deep impact of both compounds on ERα-modulated genes, including the down-regulation of ERα itself. On the other hand, functional analysis showed that commonly affected transcripts are involved in different cellular processes, such as cancer cell survival, chemoresistance and cell cycle progression. Moreover, ChIP-qPCR performed on ERα promoter highlighted ERα and DOT1L co-localization, both in PEO1 and in PEO4 cells, which was reduced after EPZ treatment, suggesting a role of this complex on receptor transcriptional activity. In addition, drug combination studies performed with EPZ and ICI showed an additive effect in cell growth inhibition. Taken together, these results suggest DOT1L as a potential therapeutic target in the treatment of OC.

2019-11-04 | E-MTAB-8198 | 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