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Effect of LIN28 on miRNA profile at day 5 of neurogliogenesis in vitro

ABSTRACT: Comparison of mouse P19 and P19+Lin28 cells at day 5 of retinoic acid-induced differentiation The cell lines differ in that one constitutively expresses Lin28 from a heterologous promoter.

ORGANISM(S): Mus musculus Rattus norvegicus Human gammaherpesvirus 8 JC polyomavirus Betapolyomavirus macacae Homo sapiens Murid gammaherpesvirus 4 Human immunodeficiency virus 1 Human betaherpesvirus 5 Betapolyomavirus hominis Human alphaherpesvirus 1 human gammaherpesvirus 4 Murid betaherpesvirus 1

PROVIDER: GSE19858 | GEO | 2010/02/25

SECONDARY ACCESSION(S): PRJNA122137

REPOSITORIES: GEO

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P19 cells+LIN28_RA-differentiated_Day 4

Project description:LIN28 is an RNA-binding protein expressed in many developing tissues. It can block let-7 microRNA processing and help promote pluripotency. We observe LIN28 expression in the developing neural tube, colocalizing with SOX2, suggesting a role in neural development. To better understand its normal developmental function, we investigated LIN28 activity during neurogliogenesis in vitro where the succession of neuronal to glial cell fates occurs as it does in vivo. LIN28 expression was high in undifferentiated cells, and was down-regulated rapidly upon differentiation. Constitutive LIN28 expression caused a complete block of gliogenesis and an increase in neurogenesis. LIN28 expression was compatible with neuronal differentiation and did not increase proliferation. LIN28 caused significant changes in gene expression prior to any effect on let-7, notably on Igf2. Furthermore, a mutant LIN28 that permitted let-7 accumulation was still able to completely block gliogenesis. Thus, at least two biological activities of LIN28 are genetically separable and may involve distinct mechanisms. LIN28 can differentially promote and inhibit specific fates and does not function exclusively by blocking let-7 family miRNAs. Importantly, LIN28âs role in cell fate succession in vertebrate cells is analogous to its activity as a developmental timing regulator in C. elegans. Two repeats of two cell lines induced to differentiate with retinoic acid/aggregation for four days. Control cells: unmodified mouse P19 embryonal carcinoma cells. Experimental cells: P19 cells constitutively expressing human LIN28A.

2010-03-03 | E-GEOD-19705 | biostudies-arrayexpress

Genome-wide analysis of RNAs associated with Lin28

Project description:We used immunoprecipitation to pulldown Lin28 associated RNA targets and used genome-wide high throughput deep sequencing to identified those Lin28-associated RNAs. Keywords: Lin28 IP-RNAseq

2011-01-18 | GSE23109 | GEO

Genome-wide analysis of RNAs associated with Lin28

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LIN28 binds messenger RNAs at GGAGA motifs and regulates splicing factor abundance (HTS)

Project description:LIN28 is a conserved RNA binding protein implicated in pluripotency, reprogramming and oncogenesis. Previously shown to act primarily by blocking let-7 microRNA (miRNA) biogenesis, here we elucidate distinct roles of LIN28 regulation via its direct messenger RNA (mRNA) targets. Through cross-linking and immunoprecipitation coupled with high-throughput sequencing (CLIP-seq) in human embryonic stem cells and somatic cells expressing exogenous LIN28, we have defined discrete LIN28 binding sites in a quarter of human transcripts. These sites revealed that LIN28 binds to GGAGA sequences enriched within loop structures in mRNAs, reminiscent of its interaction with let-7 miRNA precursors. Among LIN28 mRNA targets, we found evidence for LIN28 autoregulation and also direct but differing effects on the protein abundance of splicing regulators in somatic and pluripotent stem cells. Splicing-sensitive microarrays demonstrated that exogenous LIN28 expression causes widespread downstream alternative splicing changes. These findings identify important regulatory functions of LIN28 via direct mRNA interactions.

2012-09-10 | GSE39872 | GEO

LIN28 eRIP in Differentiating hESCs

Project description:Excluded-volume-effect-optimized RNA-IP (eRIP) of LIN28-associated mRNAs in H1 hESCs. H1s are induced to differentiate into either trophoblast (TE) or neural (N) lineages, and LIN28 eRIP performed at 0hr (Undifferentiated), 12hr TE, 24hr TE and 24hr N

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LIN28 binds messenger RNAs at GGAGA motifs and regulates splicing factor abundance (splicing array)

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Lin28/let-7 axis regulates the timig of cession of nephrogenesis

Project description:To study role of Lin28/let-7 axis on nephrogenesis, we profiled kidney transcriptom of LIN28 OE, let-7 KO, and their wild-type littermate control mice.

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Non-microRNA Binding Competitively Inhibits LIN28 Regulation

Project description:RNA binding protein (RBP) expression is finite. For RBPs that are vastly outnumbered by their potential target sites, a simple competition for binding can set the magnitude of post-transcriptional control. Here we show that LIN28, best known for its direct regulation of let-7 miRNA biogenesis, is also indirectly regulated by its widespread binding of non-miRNA transcripts. Approximately 99% of LIN28 binding sites are found on non-miRNA transcripts, like protein coding and ribosomal RNAs. These sites are bound specifically and strongly, but they do not appear to mediate direct post-transcriptional regulation. Instead, non-miRNA sites act to sequester LIN28 protein and effectively change its functional availability, thus impeding the regulation of let-7 in cells. Together, these data show that the binding properties of the transcriptome broadly influence the ability of an RBP to mediate changes in RNA metabolism and gene expression.

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LIN28 binds messenger RNAs at GGAGA motifs and regulates splicing factor abundance (HTS)

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Lin28 expression affect embryonic lung development

Project description:Lin28 expression affect embryonic lung development

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