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Knockdown of Tis7 in mouse erythroblasts

ABSTRACT: Knockdown of Tis7 in mouse erythroblasts

PROVIDER: PRJNA430877 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR6497130_1.fastq.gz	Fastqsanger.gz
	SRR6497130_2.fastq.gz	Fastqsanger.gz
	SRR6497140_1.fastq.gz	Fastqsanger.gz
	SRR6497140_2.fastq.gz	Fastqsanger.gz
	SRR6497131_1.fastq.gz	Fastqsanger.gz

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Knockdown of Tis7 in mouse erythroblasts

Project description:DNA-binding factor Tis7 (Ifrd1) shows increased ribosome recruitment during eIF2 phosphorylation during proteotoxic stress in erythroblasts. Here, we examine the effect of Tis7 knockdown on RNA expression at different time points after tunicamycin exposure.

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Next generation sequencing following CUX1 knockdown in erythroblasts

Project description:Anemia is a significant cause of morbidity and mortality in myeloid malignances. Cytogenetic changes are recurrent within malignant hematopoietic stem and progenitor cells, yet their role in anemia pathogenesis remains unknown. One recurrent karyotypic abnormality in myeloid neoplasms is the deletion of part or all of chromosome 7 [-7/del(7q)], which harbors the transcription factor and tumor suppressor gene, CUX1. CUX1 knockdown mouse models develop myeloid disease similar to that seen in humans, including a spontaneous, cell intrinsic, and lethal anemia that develops with age. Here, we elucidate the cellular and molecular mechanisms by which CUX1 regulates erythropoiesis. We demonstrate CUX1 knockdown mice have an aberrant stress erythropoiesis response and decreased survival after acute anemia induction. CUX1 insufficient erythroblasts undergo accelerated cell cycling and increased apoptosis. In line with these phenotypes, transcriptome profiling indicates that CUX1-knockdown elicits increased proliferation and decreased erythroid differentiation gene signatures. ATAC-sequencing demonstrated dramatic, global chromatin opening in CUX1-knockdown erythroblasts. As measured by fluorescence lifetime imaging, this increased chromatin accessibility is concomitant with a disruption in nuclear condensation, which is normally requisite in mammalian erythroblasts for nuclear eviction and terminal differentiation. Finally, we show that CUX1 mediates chromatin compaction by promoting histone deacetylation. Thus, rather than a transcriptional role, our data implicate in an epigenetic regulatory role for CUX1 in erythropoiesis. Furthermore, these results suggest therapeutic targeting of epigenetic regulators, such as histone acetyl transferases, may have clinical benefit for the anemias associated with loss of CUX1.

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Project description:To determine the transcriptional function (if any) of the presumed nuclear export protein Xpo7 or RanBP16 Murine fetal liver erythroid precursors (Ter119-negative cells) were isolated from C57Bl6 E14.5 embryos by magnetic depletion and infected with retroviruses containing shRNA constructs against Xpo7. They were then cultured in Epo-containing media (2U/mL) for 36hrs until they were fully differentiated and then sorted by FACS for GFP+ (infected) cells in order to isolate total RNA to be used for the profiling. Expression profiling in late cultured mouse erythroblasts before and after knockdown of gene Xpo7.

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Microarray gene expression analysis of mouse fetal liver erythroblasts after plek2 knockdown

Project description:The goal of this experiment is to determine the expression levels of the genes that are influenced by the downregulation of plek2. TER119 negative mouse fetal liver erythroblasts were purified and infected with control and plek2 shRNA retroviruses and cultured in Epo free medium for 24 hours followed by Epo medium for 24 hours. Microarray gene expression profile analysis was performed using an Illumina platform with a biological triplicate of the same experiment.

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RNA-Seq analysis of microRNA expression profiles in mouse primary CFU-E late erythroid progenitors and Ter119+ mature erythroblasts

Project description:Using RNA-seq technology, we quantitatively determined the expression profile of microRNAs during mouse terminal erythroid differentiation. CFU-E erythroid progenitors were isolated from E14.5 fetal liver as the Ter119, B220, Mac-1, CD3 and Gr-1 negative, C-Kit positive and 20% high CD71 population. Mature Ter119+ erythroblasts were isolated from E14.5 fetal liver as C-Kit negative and Ter119 positive population. Consistent with nuclear condensation and global gene expression shut down during terminal erythroid differentiation, we found that the majority of microRNAs are downregulated in more mature Ter119+ erythroblasts compared with CFU-E erythroid progenitors.

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