Metabolomics,Unknown,Transcriptomics,Genomics,Proteomics

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Transcription profiling of human cells transfected with a miR-34a LNA inhibitor to investigate its role in oncogene-induced senescence


ABSTRACT: To understand the role of miR-34a in oncogene-induced senescence, we tested global mRNA expression using microarrays of TIG3 TERT/ deltaB-RAF:ER cells transfected with a miR-34a LNA inhibitor or a scrambled control LNA in the presence or absence of deltaB-RAF activation.
TIG3 TERT/ deltaB-RAF:ER cells are primary human diploid fibroblasts immortalized by hTERT. deltaB-RAF is constitutively active due to truncation of the regulatory N-terminal domain and fused to the hormone-binding domain of the oestrogen receptor, which was modified to respond to 4-hydroxytamoxifen (4-OHT) but not beta-estradiol.
TIG3 TERT/ deltaB-RAF:ER cells were treated with 500nM 4-OHT or equal amounts of ethanol the day after seeding at 1E6 per 10cm plate, in three biological replicates. After 2 days of 4-OHT treatment, cells were transfected with LNA-miR-34a or a control LNA-scramble using Lipofectamine 2000 (Invitrogen). Total RNA was harvested 24 h post-transfection (3 days of 4-OHT treatment in total) using TRIzol reagent. Affymetrix microarray analysis (HG-U133 Plus 2.0 human) was performed.

ORGANISM(S): Homo sapiens

SUBMITTER: Jonas Vikesaa 

PROVIDER: E-MEXP-2241 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Signatures of RNA binding proteins globally coupled to effective microRNA target sites.

Jacobsen Anders A   Wen Jiayu J   Marks Debora S DS   Krogh Anders A  

Genome research 20100527 8


MicroRNAs (miRNAs) and small interfering RNAs (siRNAs), bound to Argonaute proteins (RISC), destabilize mRNAs through base-pairing with the mRNA. However, the gene expression changes after perturbations of these small RNAs are only partially explained by predicted miRNA/siRNA targeting. Targeting may be modulated by other mRNA sequence elements such as binding sites for the hundreds of RNA binding proteins (RNA-BPs) expressed in any cell, and this aspect has not been systematically explored. Acr  ...[more]

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