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Genome-scale CRISPR Screen Reveals Essential Biological Process Determining Human Lineage Specification

ABSTRACT: Genome-scale CRISPR Screen Reveals Essential Biological Process Determining Human Lineage Specification

PROVIDER: PRJNA547581 | ENA |

REPOSITORIES: ENA

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Genome-scale CRISPR Screen Reveals Essential Biological Process Determining Human Lineage Specification

Project description:Human pluripotent stem cells (hPSCs) have an unlimited capacity to self-renewal and generate any somatic cell types in the human body, offer a promising cell resource for disease modeling and transplantation. However, the systematic mechanism leading the conversion of hPSCs into lineage specification, which is the earliest cellular event during human embryonic development, is elusive. Here, combining a lineage-specific reporter with inducible CRISPR/Cas9 system, we conducted a genome-scale screening in hESCs for overall drawing the track of cell fate decision. We defined eight functional modules containing embryonic development, mRNA processing, metabolism process, epigenetic regulation, cell cycle and others, that participate in full to the lineage commitment.

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N6-methyladenosine mRNA Modification Is Essential for Human Pancreatic Lineage Specification [RNA-Seq]

Project description:Recent advances in pancreatic differentiation from human pluripotent stem cells (hPSCs) hold great potentials for disease modeling and regenerative medicine, and more precise control over the dynamic differentiation process is critical. N6-methyladenosine (m6A) is the most prevalent internal messenger RNA (mRNA) modification, while the roles of m6A mark in pancreatic differentiation and development remain elusive. In addition, ALKBH5 is a major mRNA m6A demethylase and its role in pancreatic differentiation has not been reported. Here, we firstly studied mRNA m6A dynamics during pancreatic differentiation from hPSCs. Next, using the CRISPR-based genome editing tool, we generated ALKBH5 knockout hPSC lines and found that ALKBH5 plays important roles in pancreatic specification. After that, we conducted a series of functional and mechanistic studies, and demonstrated that ALKBH5 modulated many important genes involved in pancreatic differentiation. Collectively, our findings identified ALKBH5 as an essential regulator of human pancreatic differentiation and highlighted that m6A modification presents a new layer of regulation at epitranscriptome levels during cell-fate specification.

2022-06-13 | GSE163945 | GEO

N6-methyladenosine mRNA Modification Is Essential for Human Pancreatic Lineage Specification [m6A-Seq]

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Project description:To improve the maturity of iPSC-cardiomyocytes, we addressed the cell type specification of iPSC-cardiomyocytes by microarray and analyzed in detail about cardiac maturation process. We compared gene expression between samples, and extracted several key genes for cardac maturation and check the function of these genes. We show that a number of genes expression correlate with cardiac maturation process, and found surface markers to discriminate differentiated cardiomyocytes from other immature cells.

2018-08-31 | GSE72611 | GEO

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