p53-specific transcriptome under starvation

Signaling trough cytoplasmic or nuclear action of p53 is a major response mechanism to cellular stresses. Here we perform transcriptomics after p53 re-expression on a CRISPR/Cas9 knock out background to reveal a distinct starvation-specific transcriptome response and novel nutrient-dependent p53 target genes. Overall design: In HepG2 cells, wild-type p53 was knocked out using CRISPR-Cas9. Cells were kept for 48 hours in normal growth medium (MC-medium change) or starvation medium (STV), 3 days after electroporation with p53-expression vector or empty vector (EV) as control.

p53通过细胞质或细胞核内的信号传导发挥功能，是细胞应对各类应激刺激的核心响应机制之一。本研究在CRISPR/Cas9介导的基因敲除（CRISPR/Cas9 knock out）背景下，对p53进行重表达后开展转录组学分析，旨在揭示特异性响应饥饿状态的独特转录组变化，以及全新的营养依赖型p53靶基因。实验整体设计：以HepG2细胞为模型，利用CRISPR-Cas9技术敲除其内源性野生型p53。在通过电穿孔转染法将p53过表达载体或空载对照载体（empty vector, EV）转入细胞后的第3天，将细胞分别置于正常生长培养基（MC-medium change）或饥饿培养基（STV）中培养48小时。