Genome-wide profiling for H2A.Z and genome accessibility using ATACseq in Uhrf1hepKO mouse livers compared to WT livers

This study was carried out to investigate potential changes in the genome-wide distribution of H2AZ and genome accessibility using ATACseq in livers where Uhrf1 is deleted in hepatocytes (Uhrf1-fl/fl;Alb:Cre) compared to controls (Uhrf1-fl/fl). ChIP was carried out to pull down H2AZ bound chromatin from nuclei isolated from s single Uhrf1-Fl/fl liver and a single (Uhrf1-fl/fl;Alb:Cre) male liver between 8-12 weeks of age. This chromatin fraction was then digested using micrococcal nuclease to release mononucleosomes. DNA fragments purified from nucleosomes were then used to generate libraries and sequenced. The same livers were used for chromatin accessibility using the ATAC-seq Protocol as described in Buenrostro et al (2015).

本研究旨在探究肝细胞特异性敲除Uhrf1的小鼠肝脏（Uhrf1-fl/fl;Alb:Cre）相较于对照组（Uhrf1-fl/fl）中，组蛋白H2AZ（H2AZ）的全基因组分布以及基因组可及性的潜在变化，实验采用转座酶可及性测序（ATAC-seq）技术。本研究通过染色质免疫共沉淀（ChIP）技术，从8~12周龄的单只Uhrf1-fl/fl雄性小鼠肝脏与单只Uhrf1-fl/fl;Alb:Cre雄性小鼠肝脏分离的细胞核中，富集结合有H2AZ的染色质；随后该染色质组分经微球菌核酸酶消化以释放单核小体，从核小体中纯化得到的DNA片段被用于构建测序文库并进行测序。同一批肝脏样本还被用于按照布恩罗斯特罗等人（2015）报道的转座酶可及性测序方案检测染色质可及性。