Project description:GATA6 is required for proper definitive endoderm formation. The mechanism of this process is poorly understood We used microarrays to identify genes whose expression is altered upon GATA6-depletion
Project description:Transcriptome analysis has uncovered a series of long noncoding RNAs (lncRNAs) transcribed during cell differentiation. Here, we uncovered lncRNA GATA6-AS1 is a functional lncRNA in definitive endoderm (DE) differentiation. We found GATA6-AS1 positively regulated the expression of endoderm key factor GATA6, which was different from previous reports in other biological contexts. Further investigation showed GATA6-AS1 interacted with SMAD2/3 and recruited SMAD2/3 to the promoter region of the GATA6 gene locus. In addition, overexpression of GATA6 was able to rescue the defect of DE differentiation due to the absence of GATA6-AS1, suggesting GATA6 was the functional target of GATA6-AS1 during endoderm differentiation. Ultimately, our study uncovers GATA6-AS1 is necessary for DE and pancreas differentiation, and reveals the detailed regulation mechanism between GATA6-AS1 and DE differentiation.
Project description: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 construct that permits exongenous GATA6 cDNA expression upon supplmentation of doxycycline. We differentiated GATA6 +/+ and GATA6-/- (with and without doxycyline) cells to definitive endoderm and analyzed the chromatin profile using ATAC-seq.
Project description: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 construct that permits exongenous GATA6 cDNA expression upon supplmentation of doxycycline. We differentiated GATA6 +/+ and GATA6-/- (with and without doxycyline) cells to definitive endoderm and analyzed the gene expression profile by RNA-seq.
Project description: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 construct that permits exongenous GATA6 cDNA expression upon supplmentation of doxycycline. We differentiated GATA6 +/+ and GATA6-/- (with and without doxycyline) cells to definitive endoderm and analyzed histone tail modification profiles using CHIP-seq.
Project description: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.
Project description:FGF Signaling is required for hepatic progenitor cell formation from endoderm. The mechanism of this process is poorly understood We used microarrays to identify genes directly regulated by FGF signaling in definitive endoderm that may be involved in hepatic specification.
Project description:Optimizing the efficiency of definitive endoderm differentiation is significant for the generation of diverse organ-like structures. In this study, we utilized saracatinib to enhance definitive endoderm differentiation in pluripotent stem cells. We found saracatinib significantly improved the definitive endoderm differentiation at low concentrations. To investigate the impact of 0.5 μM saracatinib on definitive endoderm differentiation of ESC H1 cells, we conducted RNA-seq analysis with differentiated cells with or without 0.5 μM saracatinib treatment.