RNA-seq of wild-type and ZNF217-knockout KOPN-8 cells

Here, we demonstrate that ZNF217 plays an oncogenic role in B-ALL. To investigate the underlying mechanisms and pinpoint potential downstream targets that contribute to ZNF217's oncogenic role, we conducted high-throughput RNA-seq on both wild-type and ZNF217-knockout KOPN-8 cells. Our analysis revealed that FOS is An important target of ZNF217 in B-ALL. Overall design: Single-cloned KOPN-8 cells stably expressing Cas9 were transduced with either ZNF217 sgRNA (sgZNF217 #1 or sgZNF217 #2) or a scrambled sgRNA control. Following validation of ZNF217 knockout efficiency, cells were harvested to isolate total RNA. After that, poly(A)+ mRNAs were enriched for sequencing. For each group, the samples were prepared and sequenced in biological duplicates.

本研究证实ZNF217在B细胞急性淋巴细胞白血病（B-cell Acute Lymphoblastic Leukemia，简称B-ALL）中发挥致癌作用。为探究其潜在分子机制并明确介导ZNF217致癌功能的潜在下游靶点，本研究对野生型及ZNF217基因敲除的KOPN-8细胞开展了高通量RNA测序（high-throughput RNA-seq）。分析结果显示，FOS是B-ALL中ZNF217的重要下游靶点。实验整体设计：将稳定表达Cas9的单克隆KOPN-8细胞分别转导ZNF217向导RNA（sgZNF217 #1或sgZNF217 #2）或乱序向导RNA对照。在验证ZNF217基因敲除效率后，收集细胞以提取总RNA，随后富集poly(A)+信使RNA（poly(A)+ mRNA）用于测序。每组样本均设置生物学重复两份，并行完成样本制备与测序。