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M6A modification segregates the stemness program from proliferation in ISCs [scRNA-seq]

ABSTRACT: Purpose: To monitor the different of ISCs and cell fate alteration upon Mettl3 deletion, we use single cell sequencing to examine the sequence information from individual cells with optimized next-generation sequencing (NGS) technologies, providing a single-cell resolution of cellular differences. Methods:Alive intestinal epithelial cells were sorted from WT and Mettl3-KO mice at 4 dpt. ScRNA-seq libraries were generated using the Chromium Single Cell 3’ Reagent Ki V3 (10X Genomics). The libraries were sequenced as paired-end with Illumina Novaseq 6000. Raw reads were aligned to the GRCm38/mm10 mouse genome, and Cell Ranger (v3.1.0) was used to estimate unique molecular identifiers (UMIs). Raw aligned features were loaded and processed using the Seurat package (v4.0.2). Low-quality cells were filtered if they expressed no more than 200 genes or with more than 20% of mitochondrial genes. Results: After cells with low information content and a high fraction of mitochondrial RNAs were excluded, 7865 cells (4467 WT and 3398 KO) were analyzed. Conclusions: Mettl3 deletion reduced the expression of stem cell signature genes but unchanged proliferation profile in ISCs. Thus, we conclude that Mettl3 deletion leads to loss of stemness but not proliferation, and enhanced regeneration and apoptosis.

ORGANISM(S): Mus musculus

PROVIDER: GSE186913 | GEO | 2023/04/13

REPOSITORIES: GEO

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