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Comparing the transcriptoms of Mettl14fl/fl and Mettl14 fl/fl Olig2-Cre oligodendrocyre precursor cell (OPCs) and mature oligodendrocytes using Next Generation Sequencing

ABSTRACT: Comparing the transcriptoms of Mettl14fl/fl and Mettl14 fl/fl Olig2-Cre oligodendrocyre precursor cell (OPCs) and mature oligodendrocytes using Next Generation Sequencing

PROVIDER: PRJNA511396 | ENA |

REPOSITORIES: ENA

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Comparing the transcriptoms of Mettl14fl/fl and Mettl14 fl/fl; Olig2-Cre oligodendrocyre precursor cell (OPCs) and mature oligodendrocytes using Next Generation Sequencing

Project description:The goal of the experiment is to determine the transcriptome alternation of Mettl14 gene inactivation in different oligodendrocyte lineage cells(OPCs and mature oligodendrocytes), by comparing the Mettl14 fl/fl; Olig2-Cre(mutant) and Mettl14fl/fl(control) OPC and mature oligodendrocyte transcriptoms. Method: RNA collected from Mettl14fl/fl control and Mettl14fl/fl;Olig2-Cre OPC and oligodendrocytes was collected by standard protocols, sequenced using next generation high throughput sequencing technology, and processed in triplicate to produce a transcriptome profile deliminating the variable gene expression in the mutant cells. Results: We identified 11809 transcripts present in the OPC transcriptome, 586 were significantly upregulated and 177 were significantly downregulated in the mutant cells; Among 12,542 transcripts present in mature oligodendrocytes, 1388 transcripts were upregulated and 1247 were downregulated in the mutant cells(significance: q-value<0.001; fold changes (FC) that exceed 2.0 fold (log2 |FC|>1) in expression and an expression level that exceeds two counted transcripts per million (log2 |CPM| >1)) Conclusions: Mettl14 ablation differentially alters the OPC and oligodendrocyte transcriptomes

2019-12-31 | GSE124243 | GEO

Comparing the transcriptoms of Mettl14fl/fl and oligodendrocyre precursor cell (OPCs) and mature oligodendrocytes(OL)

Project description:Comparing the transcriptoms of Mettl14fl/fl and oligodendrocyre precursor cell (OPCs) and mature oligodendrocytes(OL)

| PRJNA511395 | ENA

Comparing the m6A transcriptoms of Mettl14fl/fl and oligodendrocyre precursor cell (OPCs) and mature oligodendrocytes(OL) using m6A-SMART-seq

Project description:The goal of the experiment is to determine the m6A tagged transcripts after the Mettl14 gene inactivation in OPCs and mature oligodendrocytes using Z score > 0 . Methods: mRNA from total RNA of Mettl14fl/fl OPCs and mature oligodendrocytes was purified with Dynabeads Oligo (dT)25 (ThermoFisher; 61006). The equivalent amount of input and m6A-imunoprecipitated RNA were prepared for library generation using the SMART-seq protocol as described (Picelli et al., 2014). Three biological replicates of control OPCs and three biological replicates of control oligodendrocytes were sequenced using Illumina NextSeq 500. Adapters were trimmed from original reads using Trimmomatic and low-quality reads were removed. The remaining reads were then mapped to the mouse genome (mm10) using STAR aligner (Dobin et al., 2013). To measure the relative m6A level per gene, the ratio of m6A IP/ Input was first calculated. The Z scores were then obtained by comparing the ratios (m6A IP/ Input) to the mean of the group to reflect the relative m6A level per gene on a transcriptome-wide scale. Results: The m6A-seq analyses detected 3554 m6A marked transcripts in OPCs and 2606 m6A marked transcripts in oligodendrocytes.There are 2806 transcripts with the m6A mark in OPCs that were present but not marked in oligodendrocytes , and 1626 transcripts that possessed the m6A mark in oligodendrocytes but not in OPCs . Only 23 of the shared transcripts , showed the m6A mark in both OPCs and oligodendrocytes. Conclusions: The dynamic nature of the m6A mark in oligodendrocyte lineage cells suggests that it may play an important role in oligodendrocyte differentiation and CNS myelination.

2019-12-31 | GSE124202 | GEO

Comparing the m6A transcriptoms of Mettl14fl/fl and oligodendrocyre precursor cell (OPCs) and mature oligodendrocytes(OL) using m6A-SMART-seq

Project description:Comparing the m6A transcriptoms of Mettl14fl/fl and oligodendrocyre precursor cell (OPCs) and mature oligodendrocytes(OL) using m6A-SMART-seq

| PRJNA510982 | ENA

Rapid differentiation of hiPSCs into functional oligodendrocytes using an OLIG2 synthetic modified messenger RNARapid differentiation of hiPSCs into functional oligodendrocytes using an OLIG2 synthetic modified messenger RNA

Project description:We show that a synthetic modified messenger RNA (smRNA)-based reprogramming method that leads to the generation of transgene-free OLs has been developed. An smRNA encoding a modified form of OLIG2, a key TF in OL development, in which the serine 147 phosphorylation site is replaced with alanine, OLIG2S147A, is designed to reprogram hiPSCs into OLs. We demonstrate that repeated administration of the smRNA encoding OLIG2 S147A lead to higher and more stable protein expression. Using the single-mutant OLIG2 smRNA morphogen, we establish a 6-day smRNA transfection protocol, and glial induction lead to rapid NG2+ OL progenitor cell (OPC) generation (> 70% purity) from hiPSC-derived neural progenitor cells (NPCs). The smRNA-induced NG2+ OPCs can mature into functional OLs in vitro and promote remyelination in vivo. Proteomic analysis of OLIG2-binding proteins indicates that OLIG2 is bound by the heat shock protein 70 (HSP70) complex. The HSP70 complex is bound more strongly to OLIG2 with the modified phosphorylation site than to wild-type OLIG2.

2023-03-10 | PXD036975 | Pride

m6A-sequencing of Mettl14-conditional knock-out retinas and matched control.

Project description:Post-transcriptional m6A methylation on mRNA plays a key role in neural development. Here, we specifically depleted Mettl14 (a key component of m6A complex) in retina progenitor cells by crossing Mettl14 fl/fl mice with the Chx10-Egfp/Cre mouse line and investigated Mettl14 depletion-induced m6A alteration in developing retinas using m6A-seq.

2023-05-31 | GSE206014 | GEO

Single-cell RNA-sequencing of Mettl14 conditional knock-out retinas and matched controls

Project description:Post-transcriptional m6A methylation on mRNA plays a key role in neural development. Here, we specifically depleted Mettl14 (a key component of m6A complex) in retina progenitor cells by crossing Mettl14 fl/fl mice with the Chx10-Egfp/Cre mouse line and investigated cell type and transcriptome changes in the developing retinas using scRNA-seq.

2023-05-31 | GSE206013 | GEO

Seh1 Interacts with Olig2 and Brd7 to Promote Oligodendrocyte Differentiation and Myelination (ATAC_Seq)

Project description:Nuclear pore complex components (Nups) are involved in neural development and alterations in Nup genes are linked to human neurological diseases. However, the physiological functions of specific Nups and the underlying mechanisms involved in these processes remain elusive. Here we show that tissue-specific depletion of nucleoporin Seh1 causes dramatic myelination defects in the Central Nervous System (CNS). Seh1-deficient Oligodendrocyte Progenitor Cells (OPCs) proliferate properly, but fail to differentiate into mature oligodendrocytes, which impairs myelin production and remyelination after demyelinating injury. Genome-wide analyses show that Seh1 regulates a core myelinogenic regulatory network and depletion of Seh1 alters open chromatin configurations at its target genes. Mechanistically, Seh1 regulates OPCs differentiation by assembling Olig2 and Brd7 into a transcription complex at nuclear pores. Together, our results reveal that Seh1 is required for oligodendrocyte differentiation and myelination by promoting assembly of an Olig2-dependent transcription complex and expose nucleoporins as key players in the CNS.

2018-09-12 | GSE119815 | GEO

Seh1 Interacts with Olig2 and Brd7 to Promote Oligodendrocyte Differentiation and Myelination (ChIP-Seq)

2018-09-12 | GSE119814 | GEO

Seh1 Interacts with Olig2 and Brd7 to Promote Oligodendrocyte Differentiation and Myelination (RNA-Seq)

2018-09-12 | GSE119812 | GEO