EWS::FLI cooperatively binds at GGAA microsatellites via DBD-a4 helix in TTC-466 [CTCF_CnT]

Multi-omics study of DBD-a4 helix of EWS::FLI to understand the underlying mechanism of transcriptional regulation in Ewing sarcoma cell line: TTC-466 In TTC-466 cells, we knock-down endogenous EWSR1::ERG with shRNA (KD) and rescued with FLAG tagged DBD and DBD+ constructs. Cells are then harvested for RNA-Seq (3 bio reps), CUT&Tag (3 bio reps for CTCF, 4 bio reps for H3K27ac, 4 bio reps for FLAG), and Micro-C libraries (2 bio reps).

针对EWS::FLI的DNA结合结构域α4螺旋的多组学研究，旨在解析尤因肉瘤细胞系TTC-466中转录调控的潜在机制。在TTC-466细胞中，我们通过短发夹RNA（short hairpin RNA，shRNA）敲低内源性EWSR1::ERG融合基因（KD组），并分别以带有FLAG标签的DBD及DBD+构建体开展拯救实验。随后收集细胞，进行RNA测序（RNA-Seq，3次生物学重复）、CUT&Tag实验（针对CTCF的实验设置3次生物学重复、针对H3K27ac的实验设置4次生物学重复、针对FLAG标签的实验设置4次生物学重复）以及Micro-C文库构建（2次生物学重复）