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Analysing the gene expression between wildtype and Nxt1 trans-heterozygotes in larval carcasses

ABSTRACT: We used larval carcasses of both wildtype and Nxt1 trans-heterozygotes for RNA paired-end sequencing and obtained a read depth between 21M to 32M reads across all samples. A total of 1821 genes were down-regulated and 1339 genes were up-regulated. A python script extracting intron data of genes from the FlyMine database identified a trend where down-regulated genes had a higher total intron length and had more introns. Up-regulated genes had a lower total intron length and less introns when compared to the non-differentially expressed genes. Furthermore, down-regulated genes showed that the majority of genes are involved in the production of circular RNAs

ORGANISM(S): Drosophila melanogaster

PROVIDER: GSE125776 | GEO | 2019/01/29

REPOSITORIES: GEO

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The transcriptome of larval carcasses from both wildtype and Nxt1 trans-heterozygotes

Project description:The transcriptome of larval carcasses from both wildtype and Nxt1 trans-heterozygotes were assessed by RNA-Seq. A total of 1821 genes were down-regulated and 1339 genes were up-regulated. Down-regulated genes had a higher total intron length and had more introns, while up-regulated genes had a lower total intron length and less introns when compared to the non-differentially expressed genes. The majority of down-regulated genes were involved in the production of circular RNAs (circRNAS). We further investigated circRNA expression in larval carcasses from both wildtype and Nxt1 trans-heterozygotes by sequencing total RNA after RNAseR treatment. RNAseR digests linear RNA and thus enriches for circRNAs. Mock samples without RNAseR treatment were also sequenced. We found that circRNA products of Nxt1-responsive genes are reduced in Nxt1 mutants, although the nascent transcripts of these genes are produced at normal levels.

2019-08-09 | GSE135591 | GEO

Analysing the gene expression between wildtype and Nxt1 trans-heterozygotes in whole larvae

Project description:We used whole larvae of both wildtype and Nxt1 trans-heterozygotes for RNA paired-end sequencing and obtained a read depth between 6.4M to 11.1M reads across all samples. In wandering larvae, stationary larvae and white prepupae, a total of 435, 855, and 611 genes were differentially expressed. A high percentage of down-regulated genes were highly expressed/testis-specific in the Drosophila testis.

2019-01-29 | GSE125781 | GEO

Profiling lariat intermediates reveals genetic determinants of early and late co-transcriptional splicing

Project description:Long introns with short exons in vertebrate genes are thought to require spliceosome assembly across exons (exon definition), rather than introns, thereby requiring transcription of an exon to splice an upstream intron. Here, we developed CoLa-seq (co-transcriptional lariat sequencing) to investigate the timing and determinants of co-transcriptional splicing genome wide. Unexpectedly, 90% of all introns, including long introns, can splice before transcription of a downstream exon, indicating that exon definition is not obligatory for most human introns. Still, splicing timing varies dramatically across introns, and various genetic elements determine this variation. Strong U2AF2 binding to the polypyrimidine tract predicts early splicing, explaining exon definition-independent splicing. Together, our findings question the essentiality of exon definition and reveal features beyond intron and exon length that are determinative for splicing timing.

2022-11-26 | GSE204834 | GEO

Co-transcriptional splicing efficiencies differ within genes and between cell types

Project description:Pre-mRNA splicing is carried out by the spliceosome and involves splice site recognition, removal of introns, and ligation of exons. Components of the spliceosome have been shown to interact with the elongating RNA polymerase II (RNAPII), which is thought to allow splicing to occur concurrently with transcription. However, little is known about the regulation and efficiency of cotranscriptional splicing in human cells. In this study, we used Bru-seq and BruChase-seq to determine the cotranscriptional splicing efficiencies of 17,000 introns expressed across six human cell lines. We found that less than half of all introns across these six cell lines were cotranscriptionally spliced. Splicing efficiencies for individual introns showed variations across cell lines, suggesting that splicing may be regulated in a cell type–specific manner. Moreover, the splicing efficiency of introns varied within genes. The efficiency of cotranscriptional splicing did not correlate with gene length, intron position, splice site strengths, or the intron/neighboring exons GC content. However, we identified binding signals from multiple RNA binding proteins (RBPs) that correlated with splicing efficiency, including core spliceosomal machinery components—such as SF3B4, U2AF1, and U2AF2 showing higher binding signals in poorly spliced introns. In addition, multiple RBPs, such as BUD13, PUM1, and SND1, showed preferential binding in exons that flank introns with high splicing efficiencies. The nascent RNA splicing patterns presented here across multiple cell types add to our understanding of the complexity in RNA splicing, wherein RNA-binding proteins may play important roles in determining splicing outcomes in a cell type– and intron-specific manner.

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Gene expression profiling of MRC-5 cells treated with 100 ug/mL regular-length pristine SWCNTs for 72 h

Project description:A total of 30 genes were up-regulated and 19 genes were down-regulated > 1.5-fold following 100 ug/mL regular-length pristine SWCNTs treatment

2016-11-23 | GSE89375 | GEO

pre-mRNA splicing order is defined and maintains splicing fidelity across multi-intronic transcripts [II]

Project description:Combinatorially, intron excision within a given nascent transcript could proceed down any of thousands of paths, each of which would expose different dynamic landscapes of cis-elements and contribute to alternative splicing. In this study, we found that post-transcriptional multi-intron splicing order in human cells is largely predetermined, with most genes spliced in one or a few predominant orders. Strikingly, these orders were conserved across cell types and stages of motor neuron differentiation. Introns flanking alternatively spliced exons were frequently excised last, after their neighboring introns. Perturbations to the spliceosomal U2 snRNA altered the preferred splicing order of many genes, and these alterations were associated with the retention of other introns in the same transcript. In one gene, early removal of specific introns was sufficient to induce delayed excision of three proximal introns, and this delay was caused by two distinct cis-regulatory mechanisms. Together, our results demonstrate that multi-intron splicing order in human cells is predetermined, is influenced by a component of the spliceosome and ensures splicing fidelity across long pre-mRNAs.

2023-05-14 | GSE208259 | GEO

pre-mRNA splicing order is defined and maintains splicing fidelity across multi-intronic transcripts [I]

2023-05-14 | GSE208197 | GEO

Nascent-Seq Indicates Widespread Cotranscriptional pre-mRNA Splicing in Drosophila

Project description:To determine the prevalence of cotranscriptional splicing in Drosophila, we sequenced nascent RNA transcripts from Drosophila S2 cells as well as from Drosophila heads. 87% of introns assayed manifest more than 50% cotranscriptional splicing. The remaining 13% are cotranscriptionally spliced poorly, or slowly, with ~3% being almost completely retained in nascent pre-mRNA. Although individual introns showed slight but statistically significant differences in splicing efficiency, similar global levels of splicing were seen from both sources. Importantly, introns with low cotranscriptional splicing efficiencies are present in the same primary transcript with efficiently spliced introns, indicating that splicing is intron-specific. The analysis also indicates that cotranscriptional splicing is less efficient for first introns, longer introns and introns annotated as alternative. FinallyFinally, S2 cells expressing the slow RpII215C4 mutant manifest substantially less intron retention than wild-type S2 cells. Examination of Total pA and Nascent RNA from 2 different cell populations and isolated fly heads.

2011-12-01 | E-GEOD-32950 | biostudies-arrayexpress

Global co-transcriptional splicing in Arabidopsis and the correlation with splicing regulation in mature RNAs

Project description:RNA splicing and spliceosome assembly in eukaryotes occur mainly during transcription. However, co-transcriptional splicing has not yet been explored in plants. Here, we showed that in Arabidopsis thaliana, nearly all introns undergo co-transcriptional splicing and this occurs with higher efficiency for introns in protein-coding genes than for noncoding RNAs. Co-transcriptional splicing efficiency correlates with density of 19 epigenetic modifications. Total intron number and intron position are two predominant sequence features that correlate with co-transcriptional splicing efficiency, and introns with alternative 5′ or 3′ splice sites are less efficiently spliced. Furthermore, we found that trans-acting protein mutations lead to more introns with increased splicing defects in nascent RNAs than in mature RNAs, and that introns with increased splicing defects in mature RNAs are inefficiently spliced at the co-transcriptional level. Our results not only uncovered widespread co-transcriptional splicing in Arabidopsis, but also uncovered features that may affect or be affected by co-transcriptional splicing efficiency.

2019-11-25 | GSE126064 | GEO

Analysis of non-coding RNAs in drn1âÂ deletion strains

Project description:The turnover of introns spliced from pre-mRNA occurs first by debranching the lariat intron followed by destruction of the linear intron by other nucleases. We have identified a novel component of intron turnover, DRN1 (YGR093W). In order to identify RNAs affected by Drn1 turnover, we used a microarray containing probes specific to ~400 yeast non-coding RNAs to analyze a strain deleted for Drn1. These data implicated Drn1 in the turnover of a number of introns spliced from ribosomal-protein genes. Two-color experiment with biological replicate and dye swap.

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