Phase separation of OCT4 controls TAD reorganization to promote cell fate transitions

Protocol to alter a protein’s phase separation capacity to control cell fate transitions

“…OCT4 droplets can fuse, and can fast recover after photobleaching, indicating OCT4 droplets have liquid-like behavior. These claims are also validated by in vivo study showing that OCT4 can form phase-separated condensates in vivo, which can fast recover after photobleaching ( Wang et al, 2021 ). Both acidic mutation and residue deletion can disrupt OCT4 condensates in vivo and OCT4 droplets in vitro, demonstrating IDR perturbations can destroy the phase separation capacity of proteins.…”

“…Both acidic mutation and residue deletion can disrupt OCT4 condensates in vivo and OCT4 droplets in vitro, demonstrating IDR perturbations can destroy the phase separation capacity of proteins. Furthermore, the disabled phase separation ability of OCT4 mutants can be rescued by fusing an IDR of FUS or hnRNPA ( Wang et al, 2021 ). These results support that our approach can effectively control phase-separated capacity of proteins.…”

“…The major challenge is how to effectively predict mutation residues. For complete details on the use and execution of this protocol, please refer to Wang et al (2021) .…”

Human pluripotent stem cell-derived brain organoids as in vitro models for studying neural disorders and cancer

“…hPSC-derived 3D organoids are easily accessible and handled, and have more closely recapitulated the relative complicated cytoarchitecture and microenvironment of human organs. Therefore, many of them have been generated to study tumorigenesis and personalized therapy, including glioblastoma (GBM), pancreatic ductal adenocarcinoma (PDAC), lung adenocarcinomas, retinoblastoma, and colorectal cancer organoids [ 115 – 120 ]. Using this model, the researchers have uncovered new mechanistic insights into the tumor development, which are difficult to be learned from other models.…”

“…Recently, two separate groups from Huang et al and Breunig et al have generated the hPSC-based pancreatic duct-like and acinus-like organoids, which have recapitulated the properties of neonatal exocrine pancreas. Using these models introducing the PDAC-associated oncogene mutations, they revealed that GNAS R201C and KRAS G12D have lineage-dependent effects on PDAC formation in vitro and in vivo, GNAS R201C mutated in ductal organoids induced cystic growth more effectively than acinar organoids, whereas KRAS G12D expressed in acinar organoids is more effective to induce acinar-to-ductal metaplasia-like changes and model PDAC in vivo [ 119 , 120 ]. This study highlighted the advantages of hPSC-derived organoids in modeling tumor progression of heterogeneity.…”