Genome-wide maps of REST and its cofactors in mouse E14 cells. Mus musculus

We report the application of single-molecule-based sequencing technology for REST and its cofactors genome wide binding sites in E14 cells.We then combine these binding sirtes with REST regulating gene profiling, to understand REST binding and regulation in E14 cells. Overall design: Examination of REST and 5 cofactors(RCOR1, RCOR2,RCOR3,SIN3A,SIN3B) in E14 cells, REST and SIN3A endogenous antibody were used for ChIP experiment. The stable E14 cells expressing low level exogenous RCOR1, RCOR2, RCOR3,and SIN3B with V5 tag were used for ChIP experiment with V5 antibody to obtain individual ChIP DNA.

本研究报道了基于单分子测序技术在E14细胞中检测REST及其辅因子全基因组结合位点的应用。随后，我们将这些结合位点与REST调控的基因表达谱相结合，以解析E14细胞中REST的结合模式与调控机制。整体实验设计：在E14细胞中检测REST及其5种辅因子（RCOR1、RCOR2、RCOR3、SIN3A、SIN3B）的结合情况，其中使用内源性REST与SIN3A抗体开展染色质免疫共沉淀（ChIP）实验；另外，构建稳定低表达带V5标签的外源RCOR1、RCOR2、RCOR3及SIN3B的E14细胞系，使用V5抗体进行ChIP实验以获取各自的ChIP DNA样本。