全球分析鉴定 p53 的直接转录靶标。
Global Analyses to Identify Direct Transcriptional Targets of p53.
机构信息
Department of Pharmacology, University of Colorado Anschutz Medical Campus, Aurora, CO, USA.
Linda Crnic Institute for Down Syndrome, School of Medicine, University of Colorado Anschutz Medical Campus, Aurora, CO, USA.
出版信息
Methods Mol Biol. 2021;2267:19-56. doi: 10.1007/978-1-0716-1217-0_3.
The transcription factor p53 controls a gene expression program with pleiotropic effects on cell biology including cell cycle arrest and apoptosis. Identifying direct p53 target genes within this network and determining how they influence cell fate decisions downstream of p53 activation is a prerequisite for designing therapeutic approaches that target p53 to effectively kill cancer cells. Here we describe a comprehensive multi-omics approach for identifying genes that are direct transcriptional targets of p53. We provide detailed procedures for measuring global RNA polymerase activity, defining p53 binding sites across the genome, and quantifying changes in steady-state mRNA in response to p53 activation.
转录因子 p53 控制着一个具有多种细胞生物学效应的基因表达程序，包括细胞周期停滞和细胞凋亡。在这个网络中识别直接的 p53 靶基因，并确定它们如何影响 p53 激活下游的细胞命运决定，是设计靶向 p53 以有效杀死癌细胞的治疗方法的前提。在这里，我们描述了一种全面的多组学方法，用于鉴定 p53 的直接转录靶基因。我们提供了详细的程序来测量全基因组 RNA 聚合酶活性、定义 p53 结合位点，并定量分析 p53 激活后稳态 mRNA 的变化。
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