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Effect of ablation of CEACAM1 on gene expression in human regulatory T cells

ABSTRACT: To study the contribution of CEACAM1 in human Tregs, CEACAM1 was knocked out in ex-vivo expanded human Tregs by Crispr/Cas9. Tregs transfected with scramble gRNAs were used as controls. After electroporation, scramble or CEACAM1KO Tregs were subcultured with IL-2 for 7 days and RNA was isolated for genome-wide RNA-seq. Gene expression profiling analysis was performed using data obtained from RNA-seq of two replicates of CEACAM1KO and control cells.

ORGANISM(S): Homo sapiens

PROVIDER: GSE259322 | GEO | 2025/07/23

REPOSITORIES: GEO

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Project description:Cleft palate results from a mixture of genetic and environmental factors and occurs when the bilateral palatal shelves fail to fuse. The objective of this study was to search for new genes involved in mouse palate formation. Gene expression of murine embryonic palatal tissue was analyzed at the various developmental stages before, during, and after palate fusion using GeneChip? microarrays. Ceacam1 was one of the highly up-regulated genes during and after fusion in palate formation, and this was confirmed by quantitative real-time PCR. Immunohistochemical staining showed that CEACAM1 was expressed at a very low level in palatal epithelium before fusion, but highly expressed in the midline of the palate during and after fusion. To investigate the developmental role of CEACAM1, function-blocking antibody was added to embryonic mouse palate in organ culture. Palatal fusion was inhibited by this function-blocking antibody. To investigate the subsequent developmental role of CEACAM1, we characterized Ceacam1-deficient (Ceacam1-/-) mice. Epithelial cells persisted abnormally at the midline of the embryonic palate even on day E16.0, and palatal fusion was delayed in Ceacam1-/- mice. TGF?3 expression, apoptosis, and cell proliferation in palatal epithelium were not effected in the palate of Ceacam1-/-mice. CEACAM1 expression was down-regulated in Tgfb3-/- palate. However, exogenous TGF?3 did not induce CEACAM1 expression. These results suggest that CEACAM1 has roles in both the initiation of palate formation via epithelial cell adhesion and TGF signaling has some indirect effect on CEACAM1. Global gene expression profiling of palatal processes before, during and after fusion of palatal shelves We used microarray to investigate the gene expression of palatal tissue during palatal development. Palatal processes were microdissected at the stages of palatal development (before, during and after fusion) for RNA extraction and hybridization on Affymetrix microarrays.

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Effect of ablation of CEACAM1 on gene expression in human regulatory T cells

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