Disruption of RNAPII transcription elongation links Oncogenic splicing factor mutations to replciation stress and targetable alterations in chromatin landscape [NTD-chIP-seq]

To investigate changes in RNAPII distribution in response to SF3B1 mutations, we peformed ChIP-seq using antibody against N-terminal domain of RNAPII (Pan-Pol2) in K562 cells (wild type and SF3B1K700E mutant) at 4 days after mutant allele expression We then performed coverage plot analysis using data obtained from ChIP-seq to investigate RNAPII distribution changes ChIP-seq against NTD-RNAPII in K562 (wild type vs SF3B1K700E mutant) cells at 4 days, post doxycycline induction

为探究SF3B1突变对RNA聚合酶II（RNA Polymerase II，RNAPII）分布的影响，我们在突变等位基因表达4天后，使用靶向RNAPII N端结构域的抗体（Pan-Pol2），对野生型及SF3B1K700E突变型K562细胞开展染色质免疫共沉淀测序（Chromatin Immunoprecipitation sequencing，ChIP-seq）实验。随后，我们利用本次ChIP-seq获取的数据进行覆盖度绘图分析，以解析RNAPII的分布变化；同时，我们还在多西环素（doxycycline）诱导4天后，针对野生型与SF3B1K700E突变型K562细胞开展了靶向NTD-RNAPII的ChIP-seq实验。