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Loss of Tet proteins impairs mouse embryonic stem cells differentiation toward cardiomyocytes progenitors

ABSTRACT: DNA methylation mediated epigenetic regulation plays a critical role in regulating cardiomyocytes (CM) differentiation. Tet protein mediated DNA methylation oxidation have been reported to play an important role in regulating embryonic stem cell (ESC) differentiation toward different lineages. In our study, we utilized CRISPR/Cas9 based genome editing method to generated Tet single, double and triple deficient mouse ESC (mESC) expressing emGFP under NKX2.5 promoter and differentiated these cells toward CM progenitors. By modulating emGFP population as cardiac progenitor cells, we found that deletion of Tet1 and Tet2 significantly impairs mESC differentiation toward CM progenitors. Further single-cell RNA-seq analysis in differentiated control and Tet-triple knockout (TKO) mESC reveals that Tet deletion resulted in accumulation of mesoderm progenitors and impairs CM differentiation. Overexpression the catalytic domain of Tet1 could rescue the development defects in Tet-TKO mESC and restore NKX2.5 gene expression. In addition, loci-specific dCas9-Tet1CD mediated epigenome editing at Hand1 loci confirmed its directly transcriptional regulation during CM differentiation. Overall, out studies suggested that Tet protein mediated epigenomic remodeling is a genome-wide event and is essential to maintain proper transcription network during mESC differentiation toward CM progenitors.

ORGANISM(S): Mus musculus

PROVIDER: GSE168061 | GEO | 2023/02/08

REPOSITORIES: GEO

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Loss of Tet enzymes compromises proper differentiation of embryonic stem cells

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