GATA6 defines endoderm fate by controlling chromatin accessibility during differentiation of human induced pluripotent stem cells [TF_ChIP-seq]

Investigation of the role played by GATA6 in establishing the definitive endoderm chromatin accessbility profile. We used pluripotent stem cells as a model of early development. We derived GATA6-/- pluripotent cells with an inducible GATA6 or FOXA2 construct that permits exongenous GATA6 or FOXA2 cDNA expression upon supplementation of doxycycline. We differentiated GATA6+/+ and GATA6-/- (with and without doxycyline) cells to definitive endoderm and analyzed transcription factor binding profiles using CHIP-seq. Overall design: Examination of GATA6, EOMES and FOXA2 binding profiles during definitive endoderm formation in the absence and presence of GATA6.

本研究旨在探究GATA6在确立定形内胚层（definitive endoderm）染色质可及性图谱中所发挥的作用。我们以多能干细胞（pluripotent stem cells）作为早期发育的研究模型，构建了携带诱导型GATA6或FOXA2表达载体的GATA6基因纯合敲除（GATA6-/-）多能干细胞，该载体可在添加多西环素（doxycycline）后启动外源GATA6或FOXA2的cDNA表达。随后我们将GATA6基因野生型（GATA6+/+）以及GATA6基因纯合敲除（经多西环素处理与未经多西环素处理）的细胞诱导分化为定形内胚层，并通过染色质免疫共沉淀测序（ChIP-seq）分析转录因子结合图谱。实验整体设计：探究在GATA6存在与缺失的条件下，定形内胚层形成过程中GATA6、EOMES与FOXA2的结合图谱。