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RNA-seq of human foreskin fibroblast cells lacking RB, p130, and p107 treated with doxorubicin.

ABSTRACT: We used human foreskin fibroblast cells with CRISPR-Cas9 mediated knockout of RB1 and p130 (RBL2) (sgP130, sgRB1+sg130), knocked down p107 using siRNA and measured gene expression changes after doxorubicin treatmnet (24 hr, 350 nM).

ORGANISM(S): Homo sapiens

PROVIDER: GSE135842 | GEO | 2019/10/10

REPOSITORIES: GEO

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RNA-seq of human foreskin fibroblast cells lacking RB and/or p130 after doxorubicin treatment

Project description:We used human foreskin fibroblast cells with CRISPR-Cas9 mediated knockout of RB1 and p130 (RBL2) (Control, sgRB1, sgP130, sgRB1+sg130) and measured gene expression changes after doxorubicin treatmnet (24 hr, 350 nM).

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RNA-seq of human foreskin fibroblast cells lacking RB, p130, and p107 treated with doxorubicin.

Project description:RNA-seq of human foreskin fibroblast cells lacking RB, p130, and p107 treated with doxorubicin.

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Transcription profiling of mouse epidermis from Rb(F19/F19)K14cre;Rbl2-/- (pRb-;p130-) mutants to analyse functional redundancies between retinoblastoma family members

Project description:The epidermal-specific ablation of Rb gene leads to increased proliferation, aberrant differentiation, and the disengagement of these processes in vivo and in vitro. These differences in phenotype are more severe with the loss of p107, demonstrating the functional compensation between pRb and p107. As p107 and p130 also exert overlapping functions in epidermis, we have generated Rb(F19/F19)K14cre;Rbl2-/- (pRb-;p130-) mice to analyze possible functional redundancies between pRb and p130. The epidermal phenotype was very similar between pRb- and pRb-;p130- mice, suggesting that pRb and p130 activities are not redundant in epidermis. Importantly, we can correlate the proliferation differences with specific changes in gene expression between pRb-, pRb-;p107- and pRb-;p130- primary keratinocytes using microarray analysis, and explain the phenotypes in the context of altered E2F expression and functionality. Our findings support a model in which the distinct retinoblastoma family members, in conjunction with E2F members, play a central role in regulating epidermal homeostasis through specific or overlapping activities. Experiment Overall Design: Total RNA from pooled cells from each genotype were analyzed, per duplicate (pRb-/-;p107-/-, or pRb-/-;p130-/-) or triplicate (wild type, or pRb-/-), in mouse microarrays. Comparison was performed between the 4 different genotypes.

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Gene expression profiles after RNAi-mediated suppression of RB, p107 or p130 in growing, quiescent or ras-induced senescent IMR90 cells

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Transcription profiling of human IMR90 cells after RNAi suppression of RB, p107 or p130 using in growing, quiescent or ras-induced senescent IMR90 cells

Project description:The action of RB as a tumor suppressor has been difficult to define, in part, due to the redundancy of the related proteins p107 and p130. By coupling advanced RNAi technology to suppress RB, p107 or p130 with a genome wide analysis of gene expression in growing, quiescent or ras-senescent cells, we identified a unique and specific activity of RB in repressing DNA replication as cells exit the cell cycle into senescence, a tumor suppressive program. Experiment Overall Design: Expression profiles of IMR90 cells before and after RNAi-mediated supppression of RB, p107 or p130 in growing, quiescent or ras-induced senescent conditions. RNA was extracted from growing, low serum (0.1% FBS), confluent, or ras-senescent cells.

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Genome-wide analysis of RB and p130 binding in human diploid fibroblasts

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RNA-seq of human foreskin fibroblast cells lacking RB and/or p130 after doxorubicin treatment

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Genome-wide analysis of RB and p130 binding in human diploid fibroblasts

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Project description: Not available

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