Nuclear receptor corepressor (NCOR) and HDAC3 in hypothalamus [RNA-Seq]. Mus musculus

We determined genomic binding of HDAC3 in mouse hypothalamus by ChIP-seq, and identified target genes of the NCOR/HDAC3 complex in hypothalamus of NS-DADm (mutated deacetylase activation domain in NCORs) mice by RNA-seq. Overall design: Hypothalamus from 7-8 mice were pooled for one ChIP. Multiple independent ChIP samples and Input samples were pooled to generate 1 ChIP library and 1 input library followed by sequencing. For RNA-seq, total RNAs from hypothalamus of 3 WT mice and 3 NS-DADm mice were separately prepared to generate 6 libraries, followed by sequencing

本研究通过染色质免疫共沉淀测序（ChIP-seq），解析了组蛋白去乙酰化酶3（HDAC3）在小鼠下丘脑的全基因组结合特征；并借助RNA测序（RNA-seq），在NS-DADm小鼠（其核受体共抑制因子NCOR的去乙酰化酶激活结构域发生突变）的下丘脑中，鉴定得到NCOR/HDAC3复合物的靶基因。总体实验设计：将7-8只小鼠的下丘脑组织混合以完成单次ChIP实验；将多份独立的ChIP样本与Input对照样本分别混匀，构建1个ChIP测序文库与1个Input对照文库，随后开展测序。对于RNA-seq实验，分别从3只野生型（Wild Type, WT）小鼠与3只NS-DADm小鼠的下丘脑中提取总RNA，构建6个测序文库后进行测序。