Gcn5 and PCAF negatively regulate interferon β production through HAT-independent inhibition of TBK1 [RNA-Seq]. Mus musculus

Gcn5/PCAF double knockout (dKO) leads to loss of the global H3K9ac. RNA-Seq was performed to define the changes of gene expression in response to Gcn5/PCAF deletion and H3K9ac loss Overall design: PCAF-/-;Gcn5f/D MEFs were infected with retroviral Cre to delete Gcn5 to generate Gcn5/PCAF dKO cells, followed by RNA-Seq analysis using spike-in RNA as controls

Gcn5/PCAF双基因敲除（double knockout, dKO）可引发全局H3K9乙酰化修饰（H3K9ac）的缺失。为明确Gcn5/PCAF缺失及H3K9ac丢失所介导的基因表达变化，本研究实施了RNA测序（RNA-Seq）分析。实验设计概述：将PCAF基因敲除（PCAF-/-）且Gcn5为条件性敲除（Gcn5f/D）的小鼠胚胎成纤维细胞（MEFs）用携带Cre重组酶的逆转录病毒感染，以敲除Gcn5从而构建Gcn5/PCAF双基因敲除细胞，随后以掺入式内参RNA（spike-in RNA）作为对照开展RNA测序分析。