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RNA Sequencing Facilitates Quantitative Analysis of differentially expressed genes after PTBP1 knockdown in human erythroid cells

ABSTRACT: RNA Sequencing Facilitates Quantitative Analysis of differentially expressed genes after PTBP1 knockdown in human erythroid cells

PROVIDER: PRJNA417253 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6260751_1.fastq.gz	Fastqsanger.gz
	SRR6260751_2.fastq.gz	Fastqsanger.gz
	SRR6260752_1.fastq.gz	Fastqsanger.gz
	SRR6260752_2.fastq.gz	Fastqsanger.gz
	SRR6260753_1.fastq.gz	Fastqsanger.gz

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RNA Sequencing Facilitates Quantitative Analysis of differentially expressed genes after PTBP1 knockdown in human erythroid cells

Project description:Purpose: The goals of this study are to determine possible downstream targets of PTBP1 on day8 of erythroid differentiation with or without PTBP1 knockdown. Methods:gene expression profiling with or without PTBP1 knockdown in differentiated erythroblasts at day8 were generated by deep sequencing, induplicate, using Illumina GAIIx.The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays. Conclusions: heme metabolism genes were down-regulated after PTBP1 knockdown during erythroid differentiation

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RNA Sequencing Facilitates Quantitative Analysis of differentially expressed genes after UCA1 knockdown in human erythroid cells

Project description:RNA Sequencing Facilitates Quantitative Analysis of differentially expressed genes after UCA1 knockdown in human erythroid cells

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RNA Sequencing Facilitates Quantitative Analysis of differentially expressed genes during human erythroipoiesis

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Proteome changes in PTBP1 deficient hematopoietic cells

Project description:Ribosomopathies constitute a range of disabling conditions associated with defective protein synthesis mainly affecting hematopoietic stem cells (HSCs) and erythroid development. Here we demonstrate that deletion of Polypyrimidine Tract Binding Protein 1 (PTBP1) in the hematopoietic compartment led to the development of a ribosomopathy-like condition. Specifically, loss of PTBP1 was associated with a decrease in HSC self-renewal, erythroid differentiation and protein synthesis. Consistent with its function as a splicing regulator, PTBP1 deficiency led to splicing defects in hundreds of genes and we demonstrate that the up-regulation of a specific isoform of CDC42 could partly mimic the protein synthesis defect associated with loss of PTBP1. Furthermore, PTBP1 deficiency was associated with a marked defect in ribosome biogenesis and a selective reduction in the translation of mRNAs encoding ribosomal proteins. Collectively, this work identifies PTBP1 as a key integrator of ribosomal functions and highlights the broad functional repertoire of RNA binding proteins.

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Next Generation Sequencing Facilitates Quantitative Analysis of endothelial cells transfected with Ptbp1(wt), Ptbp1(T138A) or Ptbp1(T138E) plasmid Transcriptomes

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RNA Sequencing Facilitates Quantitative Analysis of differentially expressed genes after UCA1 knockdown in human erythroid cells

Project description:Purpose: The goals of this study are to determine possible downstream targets of UCA1 on day8 of erythroid differentiation with or without UCA1 knockdown. Methods:gene expression profiling with or without UCA1 knockdown in differentiated erythroblasts at day8 were generated by deep sequencing, induplicate, using Illumina GAIIx.The sequence reads that passed quality filters were analyzed at the transcript isoform level with two methods: Burrows–Wheeler Aligner (BWA) followed by ANOVA (ANOVA) and TopHat followed by Cufflinks. qRT–PCR validation was performed using TaqMan and SYBR Green assays. Conclusions: heme metabolism genes were down-regulated after UCA1 knockdown during erythroid differentiation

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Project description:Targeted disruption of the Runx1/ AML1 gene in mice has demonstrated that it is required for the emergence of definitive hematopoietic cells, but that it is not essential for the formation of primitive erythrocytes. These findings led to the conclusion that Runx1 is a stage-specific transcription factor acting only during definitive hematopoiesis. However, the zebrafish and Xenopus homologues of Runx1 have been shown to play roles in primitive hematopoiesis, suggesting that mouse Runx1 might also be involved in the development of primitive lineages. In order to identify differentially expressed genes in runx1-/- primitive erythroid, we carried out microarray analysis. Keywords: genetic modification

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