Project description:In order to identify novel genes regulated by p53, stable line containing tet-on inducible p53 construct was generated and used for gene expression analysis. Tet-on p53 inducible stable line was induced by doxycyclin for 24 hours before total RNA was extracted for array analysis.
Project description:In order to identify novel genes regulated by p53, stable line containing tet-on inducible p53 construct was generated and used for gene expression analysis.
Project description:Transcriptional profiling of H1299 non-small cell lung carcinoma cells transfected with either wt p53 or mut(175) p53 driven by the 5xHRE promoter (5 repeats of hypoxia-inducible factor response elements) and treated for 16 h with normoxia (21% O2) or hypoxia(<0.1% O2). 5xHRE promoter ensures that p53 expression is induced in hypoxic conditions only. Goal was to determine the transcriptional response of p53 in hypoxia and the 175 p53 mutant was used as a control as it is DNA-binding defective and transcription-incompetent mutant. Four-condition experiment: wt p53-transfected H1299 cells treated with normoxia, mut p53-transfected H1299 cells treated with normoxia, wt p53-transfected H1299 cells treated with hypoxia, mut p53-transfected H1299 cells treated with hypoxia. Biological replicates: 1 normoxic sample with wt p53, 1 normoxic sample with mut p53, 3 hypoxic samples with wt p53, 3 hypoxic samples with mut p53.
Project description:p53 is one of the most frequently mutated genes in human cancers, which could regulate multiple target genes involved in a widespread of cellular processes. The regulation of DNA methylation dynamics is critical for maintaining genome stability and proper gene expression. Recently a family of Ten-Eleven-Translocation (TET) proteins has been shown to possess the enzymatic activity to oxidize 5mC to 5hmC, and contribute to DNA demethylation. Here we explore the relationship between p53 binding, 5hmC modification and gene activation using human p53-deficient cells (H1299). H1299 Cells with and without p53
Project description:Take advantage of TP53-null H1299 human lung adenocarcinoma cells carrying an inducible wild-type p53 expression system to interrogate lncRNAs and mRNAs responsive to p53 by use of lncRNA microarray.
