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Analysis of alternative splicing in mouse tissues with differentiation, disease and development.

ABSTRACT: Two major goals for regenerative medicine are to reproducibly transform adult somatic cells into a pluripotent state and to control their differentiation into specific cell fates. These goals could be furthered by obtaining a complete picture of the RNA isoforms produced by these cells due to alternative splicing (AS) and alternative promoter selection (APS). To investigate the roles of AS and APS, reciprocal exon-exon junctions were interrogated on a genome-wide scale in differentiating mouse embryonic stem cells (ESCs), mouse myometrium throughout gestation and cardiac remodeling with disease using a prototype Affymetrix microarray. Using a custom analysis package named AltAnalyze (http://www.AltAnalyze.org), we identified a large number of putative AS and APS events, the majority of which were predicted to alter protein sequence and domain composition. Over a dozen AS and APS events were validated in mouse ESCs with differentiation to embryoid bodies.

ORGANISM(S): Mus musculus

PROVIDER: GSE17021 | GEO | 2009/07/10

SECONDARY ACCESSION(S): PRJNA117831

REPOSITORIES: GEO

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Analysis of alternative splicing in mouse tissues with differentiation, disease and development.

Project description:Two major goals for regenerative medicine are to reproducibly transform adult somatic cells into a pluripotent state and to control their differentiation into specific cell fates. These goals could be furthered by obtaining a complete picture of the RNA isoforms produced by these cells due to alternative splicing (AS) and alternative promoter selection (APS). To investigate the roles of AS and APS, reciprocal exon-exon junctions were interrogated on a genome-wide scale in differentiating mouse embryonic stem cells (ESCs), mouse myometrium throughout gestation and cardiac remodeling with disease using a prototype Affymetrix microarray. Using a custom analysis package named AltAnalyze (http://www.AltAnalyze.org), we identified a large number of putative AS and APS events, the majority of which were predicted to alter protein sequence and domain composition. Over a dozen AS and APS events were validated in mouse ESCs with differentiation to embryoid bodies. Alternative splicing analysis of mouse tissue differentiation, disease and developmental remodeling paradigms.

2009-07-10 | E-GEOD-17021 | biostudies-arrayexpress

Next generation sequencing discovers alternative splicing pattern among mouse embryonic stem cells, mouse embryonic fibroblasts and Dppa5a knockdown mouse embryonic stem cells and Dppa5a target RNAs

Project description:Purpose: The goals of this study are to reveal the alternative splicing pattern between mouse embryonic stem cells and mouse embryonic fibroblasts, to identify alternative splicing events regulated by Dppa5a and to discover the direct targets of Dppa5a.

2022-12-28 | GSE221654 | GEO

Widespread regulated alternative splicing of single codons accelerates proteome evolution

Project description:Thousands of human genes contain introns ending in NAGNAG motifs (N any nucleotide), where both NAGs can function as 3' splice sites, yielding isoforms differing by inclusion/exclusion of just three bases. However, the functional importance of NAGNAG alternative splicing is highly controversial. Using very deep RNA-Seq data from sixteen human and eight mouse tissues, we found that approximately half of alternatively spliced NAGNAGs undergo tissue-specific regulation and that regulated events have been selectively retained: alternative splicing of strongly tissue-specific NAGNAGs was ten times as likely to be conserved between species as for non-tissue-specific events. Further, alternative splicing of human NAGNAGs was associated with an order of magnitude increase in the frequency of exon length changes at orthologous mouse/rat exon boundaries, suggesting that NAGNAGs accelerate exon evolution. Together, our analyses show that NAGNAG alternative splicing constitutes a major generator of tissue-specific proteome diversity and accelerates evolution of proteins at exon-exon boundaries.

2012-01-05 | GSE30017 | GEO

Widespread regulated alternative splicing of single codons accelerates proteome evolution

2012-01-05 | E-GEOD-30017 | biostudies-arrayexpress

Cardiac precursor differentiation from hESCs by lentiviral promoter drug selection

Project description:Several of the essential core transcriptional control elements in human embryonic stem cells (ESCs) have been identified, but the production and function of alternative isoforms in self-renewal, pluripotency and tissue lineage specification remain largely unknown. We have modified the H9 ESC line to allow for drug selection of human pluripotent ESCs and cardiac progenitors. Exon-level microarray expression data from undifferentiated ESCs and day 40 cardiac precursors were used to identify differentially expressed and alternative splice isoforms during differentiation. Keywords: comparison RNA from a homogenous population of undifferentiated hESCs (REX1-neo promoter drug selection) and differentiated day 40 cardiomyocytes (alpha MHC-puro promoter drug selection) was isolated and profiled with exon-tiling arrays.

2008-10-24 | E-GEOD-13297 | biostudies-arrayexpress

Microarray validation of an alternative splicing database

Project description:Alternative splicing generates functional diversity in higher organisms through alternative first and last exons, skipped and included exons, intron retentions and alternative donor and acceptor sites. In large-scale microarray studies in human and mouse, emphasis so far has been placed on exon-skip events, leaving the prevalence and importance of other splice types largely unexplored. Using a new human splice variant database and a genome-wide microarray to probes thousands of splice events of each type, we measured differential expression of splice types across 6 pairs of diverse cell lines and validated the database annotation process. Results suggest that splicing in human is more complex than simple exon skip events, which account for a minority of splicing differences. The relative frequency of differential expression of the splice types correlates with what is found by our annotation efforts. In conclusion, alternative splicing in human cells is considerably more complex than the canonical example of the exon-skip. The complementary approaches of genome-wide annotation of alternative splicing in human and design of genome-wide splicing microarrays to measure differential splicing in biological samples provide a powerful high-throughput tool to study the role of alternative splicing in human biology. Keywords: alternative splicing

2007-12-08 | GSE9734 | GEO

Quantitative microarray profiling of alternative splicing in mouse tissues

Project description:We describe the application of a new microarray platform, which combines information from exon body and splice-junction probes, to analyze the regulation of 3126 alternative splicing events in ten mouse tissues. The details of the methods and algorithms are described in this paper: Revealing global regulatory features of mammalian alternative splicing using a quantitative microarray platform (Molecular Cell, Dec., 2004) Keywords = A new quantitative alternative splicing microarray platform

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2010-09-01 | E-GEOD-23522 | biostudies-arrayexpress

A network of DZF proteins controls alternative splicing regulation and fidelity [eCLIP-seq]

Project description:Proteins containing DZF (domain associated with zinc fingers) modules play important roles throughout gene expression, from transcription to translation. Derived from nucleotidyltransferases but lacking catalytic residues, DZF domains serve as heterodimerization surfaces between DZF protein pairs. Three DZF proteins are widely expressed in mammalian tissues, ILF2, ILF3, and ZFR, which form mutually exclusive ILF2-ILF3 and ILF2-ZFR heterodimers. Using eCLIP-Seq, we find that ZFR binds across broad intronic regions to regulate the alternative splicing of cassette and mutually exclusive exons. ZFR preferentially binds dsRNA in vitro and is enriched on introns containing conserved dsRNA elements in cells. Many splicing events are similarly altered upon depletion of any of the three DZF proteins; however, we also identify independent and opposing roles for ZFR and ILF3 in alternative splicing regulation. Along with widespread involvement in cassette exon splicing, the DZF proteins control the fidelity and regulation of over a dozen highly validated mutually exclusive splicing events. Our findings indicate that the DZF protein complexes form a complex regulatory network that leverages dsRNA binding by ILF3 and ZFR to modulate splicing regulation and fidelity.

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