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Transcriptome profiling of MED1 wild-type and knockout human pre-B ALL cell line 697 (RNA-seq)

ABSTRACT: Transcriptome profiling of MED1 wild-type and knockout human pre-B ALL cell line 697 (RNA-seq)

PROVIDER: PRJEB33746 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	ERR3456271_1.fastq.gz	Fastqsanger.gz
	ERR3456271_2.fastq.gz	Fastqsanger.gz
	ERR3456272_1.fastq.gz	Fastqsanger.gz
	ERR3456272_2.fastq.gz	Fastqsanger.gz
	ERR3456273_1.fastq.gz	Fastqsanger.gz

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Transcriptome profiling of MED1 wild-type and knockout human pre-B ALL cell line 697 (RNA-seq)

Project description:CRISPR/Cas9 system was used to generate mediator complex subunit 1 (MED1) knockout human pre-B ALL cell line 697. RNA-seq was performed to observe the effects of MED1 deletion on gene expression in 697.

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Transcriptomic effects of forced expression of FOXO3 and FOXO1 in pre-B ALL cell line 697 [exp2]

Project description:To investigate diferences in function between the related transcription factors FOXO1 and FOXO3 in pre-B acute lymphocytic leukaemia (ALL) cells we extracted RNA from triplicate 697 cultures four days after transduced with empty vector (SIN-SIEW) or constructs expressing FOXO1 or FOXO3. Approximately 30 million 70bp paired-end reads were generated per sample by illumina sequencing.

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Transcriptomic effects of forced expression of FOXO3, PRDM1⍺ and PRDM1β in pre-B ALL cell line 697 [exp1]

Project description:Lentiviral constructs expressing candidate tumoure suppressor genes were transduced into 697 cells. Of the candidates FOXO3, two isoforms of PRDM1 (⍺ and β) suppressed growth in-vitro and in-vivo. To indentify tumour suppressor mechanisms we extracted RNA from triplicate 697 cultures four days after transduced with empty vector (SIN-SIEW) or constructs expressing FOXO3, PRDM1⍺ or PRDM1β. Approximately 90 million 100bp paired-end reads were generated per sample by illumina sequencing.

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Transcriptome profiling of human pre-B ALL cell line 697 upon PBX1 or TCF3 knockdown (RNA-seq)

Project description:RNA-seq was performed to observe the gene expression changes in 697 cells following shRNA-mediated knockdown of PBX Homeobox 1 (PBX1) or Transcription Factor 3 (TCF3/E2A), each with two shRNAs. Cells treated with scrambled control shRNA were used as controls.

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Genome-wide maps of factors and gene expression in pre-B leukemic 697 line

Project description:E2A, a basic helix-loop-helix (bHLH) transcription factor, plays a crucial role in determining tissue-specific cell fate, including differentiation of B cell lineages. In 5% of childhood acute lymphoblastic leukemia (ALL), the t(1,19) chromosomal translocation specifically targets the E2A gene and produces an oncogenic E2A-PBX1 fusion protein. While previous studies have demonstrated oncogenic functions of E2A-PBX1 in cell and animal models, the E2A-PBX1-enforced cistrome, the E2A-PBX1 interactome, and related mechanisms underlying leukemogenesis remain unclear. Here, by unbiased genomic profiling approaches, we identify the direct target sites of E2A-PBX1 in t(1,19)-positive pre-B ALL cells and show that, compared to normal E2A, E2A-PBX1 preferentially binds to a subset of gene loci co-bound by RUNX1 and gene-activating machineries (p300, MED1, and H3K27 acetylation). Using biochemical analyses, we further document a direct interaction between E2A-PBX1 and RUNX1 and show that E2A-PBX1 binding to gene enhancers is dependent on RUNX1, but not the DNA-binding activity harbored within the PBX1 homeodomain of E2A-PBX1. Transcriptome analyses and cell transformation assays further establish a significant RUNX1 requirement for E2A-PBX1-mediated target gene activation and leukemogenesis. Notably, the RUNX1 locus itself is also directly activated by E2A-PBX1, indicating a multilayered interplay between E2A-PBX1 and RUNX1. Collectively, our study provides the first unbiased profiling of the E2A-PBX1 cistrome in pre-B ALL cells and reveals a previously unappreciated pathway in which E2A-PBX1 acts in concert with RUNX1 to enforce transcriptome alterations for the development of pre-B ALL.

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