Pooled CRISPR screens in human intestinal organoids [CUT&RUN]. Pooled CRISPR screens in human intestinal organoids [CUT&RUN]

We performed CRISPR screens on both a sub-library and a genome-wide scale in human intestinal organoids to discover cancer driver genes. We investigated the Wnt and the TGFB pathway and used both WT, APC-mutant and APC-TP53-mutant organoids. Overall design: To investigate how the identified SWI/SNF screening hits confer TGFB resistance, we assessed changes in binding of SMAD3 to the chromatin upon ARID1A or SMARCA4 perturbation and TGFB treatment

本研究在人肠道类器官中，分别采用亚文库与全基因组规模的实验策略开展CRISPR筛选（CRISPR Screen），旨在挖掘癌症驱动基因。本研究聚焦Wnt信号通路与转化生长因子β（TGF-β）通路，所用实验模型涵盖野生型（WT）、APC突变型以及APC-TP53双突变型肠道类器官。整体实验设计：为探究已筛选获得的SWI/SNF复合物（SWI/SNF Complex）靶点如何赋予细胞TGF-β抗性，本研究分别在ARID1A或SMARCA4基因扰动以及TGF-β处理的条件下，检测了SMAD3与染色质的结合水平变化。