PRDM16 binding in brown adipocyte precursors [ChIP-seq]

PRDM16 is an important transcriptional regulator of brown and beige adipocyte gene programs. We discovered that PRDM16 also represses type I Interferon-stimulated genes (ISGs) in brown and beige adipocytes. ChIP-seq demonstrated that PRDM16 binds proximal to many of the regulated ISGs and this correlates with reduction in the amount of the activating mark, H3K27Ac. Together this data shows PRDM16 represses ISG expression through direct binding at promoter regions to block transcription. Overall design: Immortalized adipocyte precursors derived from Prdm16 KO brown adipose were infected with control or PRDM16 expressing plasmids. ChIP-seq was performed for PRDM16 and H3K27Ac to determine PRDM16 binding in adipocyte precursors.

PRDM16是调控棕色与米色脂肪细胞基因程序的关键转录调控因子（transcriptional regulator）。本研究发现，PRDM16在棕色及米色脂肪细胞中还可抑制I型干扰素刺激基因（type I Interferon-stimulated genes, ISGs）的表达。染色质免疫共沉淀测序（ChIP-seq）结果显示，PRDM16可结合于多数受调控ISGs的近端基因组区域，且该结合与激活性组蛋白修饰标记H3K27乙酰化（H3K27Ac）的水平下降存在显著相关性。综合全部实验数据表明，PRDM16可通过直接结合靶基因启动子区域阻断转录，进而抑制ISGs的表达。
整体实验设计：将源自Prdm16敲除（Knockout, KO）棕色脂肪组织的永生化脂肪细胞前体细胞，分别感染空载对照质粒与过表达PRDM16的重组质粒。随后针对PRDM16及H3K27Ac开展ChIP-seq实验，以解析脂肪细胞前体细胞中PRDM16的结合特征。