Tbx1 binding site in whole mouse embryo at E9.5 from in vivo ChIP-seq

We generated Avidin tag inserted at 3' end of Tbx1 using CRISPR/Cas9 genomic engineering and crossed them with BirA mice from the Jackson laboratory. After established double homozygous mouse colony, total twenty embryos at E9.5 were collected for each samples. After extraction of nucleus, Tbx1 binding regions were captured with Dynabeads™ MyOne™ Streptavidin T1. Overall design: Tbx1 is T-box transcription factor which expresses in pharyngeal apparatus in mouse embryo. To capture Tbx1 binding site in vivo, nucleus from Tbx1-Avi;BirA double homozygous embryos at E9.5 were collected for ChIP-assay with streptavidin beads

我们通过CRISPR/Cas9基因组工程技术（CRISPR/Cas9 genomic engineering），在Tbx1基因的3'端插入抗生物素蛋白标签（Avidin tag），并将构建得到的工程小鼠与来自杰克逊实验室（Jackson Laboratory）的BirA小鼠进行杂交。待双纯合子小鼠种群建立完成后，我们为每个样本收集了共计20枚胚胎发育第9.5天（E9.5）的小鼠胚胎。细胞核提取完成后，使用Dynabeads™ MyOne™ Streptavidin T1磁珠捕获Tbx1结合区域。
实验整体设计：Tbx1属于T-box转录因子（T-box transcription factor），在小鼠胚胎的咽装置中表达。为了在体内捕获Tbx1的结合位点，我们收集了胚胎发育第9.5天的Tbx1-Avi;BirA双纯合子小鼠胚胎的细胞核，采用链霉亲和素磁珠开展染色质免疫沉淀试验（ChIP assay）。