RUNX1 overexpression in precursor B acute leukemia cells

Overexpression of the transcription factor RUNX1 was studied in Nalm-6 precursor B acute lymphoblastic leukemia cells. The active enhancer histone marker H3K27ac and RUNX1 were measured using ChIP-seq. The effect on gene expression was analyzed using single cell RNA-sequencing. ChIP-seq profiles were generated from Nalm-6 pre-B-ALL cell line after induced wt RUNX1 or empty vector expression for 48 h. Empty vector induction cell line was used as controls. Mnase digestion was used for DNA fragmentation. Antibodies against H3K27ac and RUNX1 compared to input. scRNA-seq samples were generated from 72 h after induction using the 10X Genomics Chromium platform.

本研究在Nalm-6前体B细胞急性淋巴细胞白血病（precursor B acute lymphoblastic leukemia）细胞中开展了转录因子RUNX1过表达的相关实验。研究通过染色质免疫共沉淀测序（Chromatin Immunoprecipitation sequencing, ChIP-seq）检测活性增强子组蛋白标记H3K27ac与RUNX1的结合谱；利用单细胞RNA测序（single cell RNA-sequencing, scRNA-seq）分析其对基因表达的影响。ChIP-seq测序数据构建自经野生型RUNX1（wt RUNX1）或空载体诱导表达48小时的Nalm-6前B急性淋巴细胞白血病细胞系，其中空载体诱导的细胞系作为对照样本。实验采用微球菌核酸酶（Micrococcal Nuclease, Mnase）酶解进行DNA片段化，以Input样本作为针对H3K27ac与RUNX1的抗体的染色质免疫共沉淀对照。单细胞RNA测序样本采集自诱导表达72小时后的细胞，测序平台采用10X Genomics Chromium平台。