Zscan4 binds nucleosomal microsatellite DNA and protects mouse two-cell embryos from DNA damage [ChIP-seq]

Genome-wide binding analysis of Zscan4 showed sequence-specific occupancy at (CA)n microsatellite repeats in 2C-like cells. Genome-wide occupancy profiling of FACT subunit SSRP1 in curaxin-treated mouse ESC demonstrated re-localization from transcription start sites to Zscan4 binding sites. We profiled SSRP1 binding sites by performing ChIP-seq using an SSRP1 antibody in control or curaxin-treated mouse ESC. There are twelve samples analyzed and two replicates for each treatment. We also identified direct targets of Zscan4 in 2C-like cells by performing ChIP-seq using a GFP antibody in 2C-like cells (GFP positive population) from the Zprom::GFP-Zscan4 reporter line and control Zprom::GFP line, and profiled genomic occupancy of endogenous Zscan4 using a Zscan4 antibody in the Zprom::GFP line (GFP positive population and control GFP negative population). There are eight samples analyzed.

对Zscan4开展全基因组结合分析后发现，其在2C样细胞中可特异性结合于(CA)ₙ微卫星重复序列位点。对经curaxin处理的小鼠胚胎干细胞（ESC）中的染色质转录辅助因子（FACT）亚基SSRP1进行全基因组结合谱分析，结果显示其结合位点从转录起始位点重新定位至Zscan4结合区域。本研究通过在对照组及curaxin处理组小鼠ESC中使用SSRP1抗体开展染色质免疫共沉淀测序（ChIP-seq），对SSRP1的结合位点进行了谱分析；本次分析共纳入12个样本，每组处理设置2次生物学重复。本研究还通过两种策略鉴定2C样细胞中Zscan4的直接靶标：其一，从Zprom::GFP-Zscan4报告细胞系与对照Zprom::GFP细胞系中分离2C样细胞（GFP阳性群体），使用绿色荧光蛋白（GFP）抗体开展ChIP-seq；其二，在Zprom::GFP细胞系中使用Zscan4抗体开展ChIP-seq，分析内源性Zscan4的基因组结合谱，分别纳入GFP阳性群体与对照GFP阴性群体。本次分析共纳入8个样本。