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Divergent transcription of lncRNA/mRNA gene pairs in embryonic stem cells

ABSTRACT: Many long non-coding RNA (lncRNA) species have been identified in mammalian cells, but the genomic origin, regulation and function of these molecules in individual cell types is poorly understood. We have generated comprehensive catalogs of lncRNA species expressed in human and murine embryonic stem cells (ESCs) and mapped their genomic origin. A surprisingly large fraction of these transcripts (>60%) originate from divergent transcription at promoters of active protein-coding genes. The divergently transcribed lncRNA/mRNA gene pairs exhibit coordinated changes in transcription when ESCs are differentiated into endoderm. A significant number of the divergently transcribed lncRNAs/mRNA pairs are conserved between human and mouse ESCs. Disruption of promoter-associated lncRNA orthologs in a zebrafish model of early development causes gross developmental defects. Our results reveal that transcription of most lncRNA genes is coordinated with transcription of protein-coding genes and that these lncRNAs have roles in early development.

ORGANISM(S): Homo sapiens

PROVIDER: GSE41009 | GEO | 2013/01/02

SECONDARY ACCESSION(S): PRJNA175529

REPOSITORIES: GEO

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Divergent transcription of lncRNA/mRNA gene pairs in embryonic stem cells

Project description:Many long non-coding RNA (lncRNA) species have been identified in mammalian cells, but the genomic origin, regulation and function of these molecules in individual cell types is poorly understood. We have generated comprehensive catalogs of lncRNA species expressed in human and murine embryonic stem cells (ESCs) and mapped their genomic origin. A surprisingly large fraction of these transcripts (>60%) originate from divergent transcription at promoters of active protein-coding genes. The divergently transcribed lncRNA/mRNA gene pairs exhibit coordinated changes in transcription when ESCs are differentiated into endoderm. A significant number of the divergently transcribed lncRNAs/mRNA pairs are conserved between human and mouse ESCs. Disruption of promoter-associated lncRNA orthologs in a zebrafish model of early development causes gross developmental defects. Our results reveal that transcription of most lncRNA genes is coordinated with transcription of protein-coding genes and that these lncRNAs have roles in early development. Analysis of genome-wide lncRNA transcription in human embryonic stem cells and early differentiation using RNA-seq, GRO-seq and ChIP-seq

2013-01-02 | E-GEOD-41009 | biostudies-arrayexpress

Evx1as and EVX1 knockouts by CRISPR

Project description:We report that long noncoding RNAs contribute to transcription and developmental process. Thousands of lncRNAs have been identified in the whole genome, and tend to located closely to protein-coding genes. To study position relationship between lncRNA and protein-coding genes, we classified all of lncRNA to several subgroups based on the genome position with their coding neighbors. XH, the head to head subgroup is associated with transcription and development in GO analysis. Here, we knockdown serveral XH lncRNA by shRNA in embryonic stem cells and induce nondirectional differnetiation by removing LIF or neural differnetiation by RA. Knockdown of XH lncRNAs led to uniform downregulation of nearby coding genes, and form regulatory circuits with its nearby coding genes to fine-tune embryonic lineage development. In addition, we also knockout one lncRNA-Evx1as and its nearby protein-coding gene-EVX1 by CRISPR, and get similar results as knockdown.We propose that XH lncRNA may function primarily as 'cis-regulators' of the expression of nearby protein-coding genes, and tend to participate in transcriptional or development regulations as their coding neighbors. All RNA-seq(s) were designed to reveal the differentially expressed genes between wild-type and XH lncRNA knockdown/knockout ESCs during differentiation.

2016-03-24 | E-GEOD-70419 | biostudies-arrayexpress

cis regulated role of lncRNA in transcriptional or developmental process (ChIRP-Seq)

2016-03-24 | E-GEOD-79541 | biostudies-arrayexpress

The long noncoding RNA expression profile of hepatocellular carcinoma identified by microarray analysis

Project description:To explore the potential involvement of lncRNAs in hepatocellular carcinoma (HCC) oncogenesis, we conducted lncRNA and mRNA profiling in 3 pairs of human HCC and adjacent normal tissue (NT) by microarray. With abundant probes accounting for 33,045 lncRNAs and 30,215 coding transcripts in our microarray, the number of lncRNAs and coding transcripts that could be detected here is 10,149 and 14,944, respectively. From the data, thousands of lncRNAs and mRNAs were found to bedifferentially expressed (Fold Change≥2.0) in HCC tissues compared with NT and identified 624 lncRNAs and 1050 mRNAs were differentially expressed in all three HCC tissues.Bioinformatic analysis (gene ontology, pathway and network analysis) was performed for further study of these differentially expressed mRNAs.By qRT-PCR analysis in nineteen pairs HCC and adjacent normal tissues, we found that eightl ncRNAs were aberrantly expressed in HCC compared with corresponding NT, which is consistent with microarray data. Additionally, change trends of seven lncRNAs were basically identical to their nearby coding genes. In this study, to explore the potential involvement of lncRNAs in hepatocellular carcinoma (HCC) oncogenesis, we conducted lncRNA and mRNA profiling in 3 pairs of human HCC and adjacent normal tissue (NT) by microarray.

2014-02-21 | E-GEOD-55191 | biostudies-arrayexpress

Comprehensive Identification of Soybean Long Non-coding RNAs Reveals a Subset of Small Peptide-Coding Transcripts

Project description:Long non-coding RNAs (lncRNAs) are defined as non-protein-coding transcripts that are at least 200 nucleotides long. They are known to play pivotal roles in regulating gene expression, especially during stress responses in plants. We used a large collection of in-house transcriptome data from various soybean (Glycine max and Glycine soja) tissues treated under different conditions to perform a comprehensive identification of soybean lncRNAs. We also retrieved publicly available soybean transcriptome data that were of sufficient quality and sequencing depth to enrich our analysis. In total, RNA-seq data of 332 samples were used for this analysis. An integrated reference-based, de novo transcript assembly was developed that identified ~69,000 lncRNA gene loci. We showed that lncRNAs are distinct from both protein-coding transcripts and genomic background noise in terms of length, number of exons, transposable element composition, and sequence conservation level across legume species. The tissue-specific and time-specific transcriptional responses of the lncRNA genes under some stress conditions may suggest their biological relevance. The transcription start sites of lncRNA gene loci tend to be close to their nearest protein-coding genes, and they may be transcriptionally related to the protein-coding genes, particularly for antisense and intronic lncRNAs. A previously unreported subset of small peptide-coding transcripts was identified from these lncRNA loci via tandem mass spectrometry, which paved the way for investigating their functional roles. Our results also highlight the current inadequacy of the bioinformatic definition of lncRNA, which excludes those lncRNA gene loci with small open reading frames (ORFs) from being regarded as protein-coding.

2020-01-09 | PXD014553 | Pride

A fine-tuned regulation of Bmp2 by Nodal/TGFβ signalling induced lncRNA-Smad7

Project description:The transforming growth factor beta (TGF-β) superfamily proteins are potent regulators of cellular development and differentiation. Long non-coding RNAs (lncRNAs) play widespread roles in spatial-temporal regulation of early development. However, the roles of lncRNAs regulated by nodal/TGF-β signaling is still elusive. Here, we showed a nodal-driven Smad induced lncRNA in mouse embryonic stem cells (mESCs), lncRNA-Smad7, which is divergently transcribed to Smad7, regulates cell fate determination through repressing Bmp2. Depletion of lncRNA-Smad7 dramatically impairs cardiomyocyte differentiation in mESCs. Moreover, LncRNA-Smad7 represses Bmp2 expression and binds at the promoter region of Bmp2. Importantly, knock-down Bmp2 rescues the defect of cardiomyocyte differentiation. Hence, we showed that lncRNA-Smad7 is antagonistic to BMP signaling in mESCs. Furthermore, lncRNA-Smad7 regulates cell fate determination between osteocytes and myocytes formation in C2C12 cells by repressing Bmp2. Thus, we provide new insights regarding the antagonistic effects between nodal/TGF-β and BMP signaling via lncRNA-Smad7.

2022-10-19 | GSE178136 | GEO

Profiles of Long Noncoding RNAs in Human Naive and Memory T Cells

Project description:We employed whole-genome RNA-sequencing to profile mRNAs and both annotated and novel long noncoding RNAs (lncRNAs) in human naive, central memory, and effector memory CD4+ T cells. Loci transcribing both lineage-specific annotated and novel lncRNA are adjacent to lineage-specific protein-coding genes in the genome. Lineage-specific novel lncRNA loci are transcribed from lineage-specific typical- and supertranscriptional enhancers and are not multiexonic, thus are more similar to enhancer RNAs. Novel enhancer-associated lncRNAs transcribed from the IFNG locus bind the transcription factor NF-κB and enhance binding of NF-κB to the IFNG genomic locus. Depletion of the annotated lncRNA, IFNG-AS1, or one IFNG enhancer-associated lncRNA abrogates IFNG expression by memory T cells, indicating these lncRNAs have biologic function.

2017-06-11 | GSE85294 | GEO

Long noncoding RNA profiling in HUVEC stimulated with IL-1β.

Project description:The purpose of this study was to identify long noncoding RNAs (lncRNAs) that are dysregulated during vascular inflammation and contribute to mRNA regulation. Previous studies have suggested that many lncRNAs interact with protein complexes to regulate their neighboring mRNAs. Therefore, we performed the Human LncRNA Expression Microarray V3.0 (Arraystar) to simultaneously profile the expression of lncRNAs and mRNAs under basal conditions and upon stimulation with IL-1β in human umbilical vein endothelial cells (HUVECs). Following, we identified neighboring IL-1β mRNA-lncRNA pairs and found them to be highly correlation in expression. This observation was particularly true when the pairs were found within the same chromatin neighborhood. We also observed mRNA-lncRNA pairs to be mainly divergent transcribed and share common regulatory elements.

2019-08-26 | GSE127990 | GEO

Deregulated Expression of Mammalian lncRNA through Loss of SPT6 Induces R-Loop Formation, Replication Stress, and Cellular Senescence

Project description:RNA polymerase II (Pol II) generates interspersed protein-coding and long noncoding (lnc) transcripts across the mammalian genome. We recently established that lncRNAs are transcribed by different Pol II CTD isoforms correlating with reduced RNA processing efficiency and increased RNA turnover. How protein-coding and lncRNA transcription units are distinguished during gene expression remains unclear. We now show that H3K36me3 chromatin is a predominant epigenetic mark for active protein-coding but not lncRNA genes. Surprisingly depletion of the Pol II associated transcription elongation factor SPT6, causes redistribution of H3K36me3 histone marks to lncRNA genes which correlates with increased lncRNA transcription. SPT6 knockdown also reduces recruitment of Integrator complex to chromatin resulting in a transcription termination defect for lncRNA genes. Notably these extended lncRNAs are both chromatin-restricted and form increased levels of RNA:DNA hybrid (R-loops) with associated DNA damage. This results in a cellular senescence phenotype and underlies the importance of restricting lncRNA expression.

2018-11-21 | GSE110028 | GEO

Long non-coding RNAs from divergent transcription of protein-coding genes

Project description:A remarkable number of long non-coding RNA (lncRNA) species have been identified in mammalian cells, but the genomic origins of these molecules in individual cell types is poorly understood. As a prerequisite to studying the transcriptional regulation of lncRNAs, we conducted a comprehensive analysis of the genomic origins of lncRNAs expressed in embryonic stem cells (ESCs). Polyadenylated RNA and total RNA depleted of ribosomal content was used for preparation of two independent sequencing libraries

2012-08-28 | E-GEOD-36799 | biostudies-arrayexpress

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