p53-specific transcriptome under starvation. 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通过胞质或核内的信号通路发挥作用，是细胞应对应激的主要响应机制。本研究在CRISPR/Cas9敲除（CRISPR/Cas9 knockout）背景下对p53进行重新表达，随后开展转录组学（transcriptomics）分析，以揭示独特的饥饿特异性转录组响应以及新的营养依赖型p53靶基因。
实验整体设计：在HepG2细胞中利用CRISPR-Cas9技术敲除野生型p53。在通过电穿孔法转染p53表达载体或空载对照载体（empty vector, EV）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。