Expression profiling in STAG2 mutant K562 cells

K562 (female chronic myelogenous leukemia) cells were CRISPR-Cas9 edited to contain STAG2 R614* mutation. STAG2 is encoded on the X-chromosome and normally mutation on one allele is sufficient for complete loss of STAG2 protein due to the wild type allele being mosaically silenced. However, we obtained both heterozygous and homozygous mutant lines that showed partial and complete loss of STAG2 protein respectively. We here examined the impact of STAG2 R614* mutation on gene expression and chromatin acessibilty. Overall design: RNA-sequencing, in triplicate, on the parental or wild type K562 cells (K562WT), two K562 cell lines that are heterozygous (STAG2heterozygous) for STAG2 R614* mutation and two K562 cell lines that are homozygous for STAG2 R614* (STAG2homozygous) mutation.

本研究通过CRISPR-Cas9基因编辑技术，将STAG2 R614*突变引入K562（女性慢性髓系白血病）细胞中。STAG2基因定位于X染色体，正常情况下，由于野生型等位基因会发生镶嵌式沉默，仅单等位基因突变即可导致STAG2蛋白完全缺失。但本研究中我们同时获得了杂合突变与纯合突变细胞系，二者分别表现出STAG2蛋白部分缺失与完全缺失。本研究旨在探究STAG2 R614*突变对基因表达及染色质可及性的影响。总体实验设计：对亲本/野生型K562细胞（K562WT）、2株携带STAG2 R614*杂合突变的K562细胞系（STAG2heterozygous）以及2株携带STAG2 R614*纯合突变的K562细胞系（STAG2homozygous）进行三次生物学重复的RNA测序。