Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Gene expression profiles in semi-continuous culture with high-cell density under normal and low temperature conditions for scFv-Fc-producer CHO cells harboring an artificial promoter system with transcriptional positive-feedback loop of transactivator

ABSTRACT: We developed an artificial gene expression system with transcriptional positive-feedback loop of transactivator to induce the high expression of target genes in CHO cells. DNA microarray analysis was conducted in semi-continuous culture with high-cell-density culture under normal and low temperature conditions of recombinant scFv-Fc-producer CHO cells (CHO/aTF_scFv-Fc2 cells).

ORGANISM(S): Cricetulus griseus

SUBMITTER: Masamichi Kamihira

PROVIDER: E-MTAB-13440 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Î³-secretase is an intra-membrane-cleaving aspartyl protease implicated in the processing of a wide range of type I membrane proteins including the Notch receptor and the amyloid-Î² precursor protein (APP). It thus regulates a diverse array of cellular and biological processes including the differentiation of neuronal embryonic stem cells, or intestinal stem cells, with the latter controlling the self-renewing of the intestinal epithelium. Indeed, proteolysis of these proteins by Î³-secretase triggers signaling cascades by releasing intracellular domains (ICDs) which, following association with adaptor proteins and nuclear translocation, modulate the transcription of different genes by binding directly to their promoters. The pronounced proliferative and regenerative effects of Notch signaling and its implication in the generation of the AÎ²-peptides, makes Î³-secretase a therapeutic target for several types of cancer and for Alzheimerâs disease. To investigate the broad effects of Î³-secretase activity onto the cellular transcriptome, Chinese hamster ovary (CHO) cells with enhanced Î³-secretase were compared to cells with abolished Î³-secretase activity via a microarray designed for a genetically close species, mouse. Our findings will potentially help to decipher the biology of Î³-secretase, including a better understanding of the roles of this enzyme in gene transcription. We compared the transcriptomes of two CHO cell lines displaying extreme differences in Î³-secretase activity. The S-1 cell line overexpressed the four components of Î³-secretase (NCT, APH1aL, PS1, and PEN2) and was characterized by a marked increase in the level of PS1 heterodimers associated with 8-fold increased Î³-secretase activity compared to untransfected controls. The other cell line consisted of wild type CHO cells incubated with DAPT, a well-known Î³-secretase inhibitor. The two cell lines were used in combination with a mouse microarray to analyze gene transcription under enhanced Î³-secretase. Two samples, S1 and DAPT treated CHO- were used in biological triplicates each.

Dataset Information

Gene expression profiles in semi-continuous culture with high-cell density under normal and low temperature conditions for scFv-Fc-producer CHO cells harboring an artificial promoter system with transcriptional positive-feedback loop of transactivator

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