SETD2 Mutations Do Not Contribute to Clonal Fitness in Response to Chemotherapy in Childhood B Acute Lymphoblastic Leukemia (WES). SETD2 Mutations Do Not Contribute to Clonal Fitness in Response to Chemotherapy in Childhood B Acute Lymphoblastic Leukemia (WES)

To investigate the impact of SETD2 mutations on clonal fitness in B actue lymphoblastic leukemia we generated cells lines with SETD KO using CRISPR/Cas9 system. We then performed phenotypic based assays as well as gene expression profiling, mutation burden, and chromtain accessibilty analysis using data obtained from RNA-seq, Whole exom sequencing following drug traetment, and ATAC-seq. Overall design: Mutation calling between pretreated and treated clone for KOPN-8 KO and WT clone. 3 clones of each cell line was subjected to treatment and sequences before and after treatment with chemotherap.

为探究SETD2突变对B细胞急性淋巴细胞白血病克隆适应性的影响，我们利用CRISPR/Cas9系统构建了SETD2敲除的细胞系。随后，我们借助RNA测序（RNA-seq）、药物处理后的全外显子组测序（Whole Exome Sequencing）及转座酶可及性测序（ATAC-seq）获得的数据，开展了基于表型的检测、基因表达谱分析、突变负荷分析与染色质可及性分析。整体实验设计：针对KOPN-8细胞系的敲除与野生型（Wild Type, WT）克隆，对预处理组与药物处理组的克隆进行突变位点识别。每个细胞系设置3个生物学重复克隆，分别接受化疗药物处理，并在处理前后完成测序。