title: Regulation of Pluripotency-related Genes and Differentiation in Mouse Embryonic Stem Cells by Direct Delivery of Cell-penetrating Peptide-conjugated CARM1 Recombinant Protein.
abstract: Coactivator-associated arginine methyltransferase 1 (CARM1) is included in the protein arginine methyltransferase (PRMT) family, which methylates histone arginine residues through posttranslational modification. It has been proposed that CARM1 may up-regulate the expression of pluripotency-related genes through the alteration of the chromatin structure. Mouse embryonic stem cells (mESCs) are pluripotent and have the ability to self-renew. The cells are mainly used to study the genetic function of novel genes, because the cells facilitate the transmission of the manipulated genes into target mice. Since the up-regulated methylation levels of histone arginine residue lead to the maintenance of pluripotency in embryos and stem cells, it may be suggested that CARM1 overexpressing mESCs elevate the expression of pluripotency-related genes in reconstituted embryos for transgenic mice and may resist the differentiation into trophectoderm (TE). We constructed a fusion protein by connecting CARM1 and 7X-arginine (R7). As a cell-penetrating peptide (CPP), can translocate CARM1 protein into mESCs. CPP-CARM1 protein was detected in the nuclei of the mESCs after a treatment of 24 hours. Accordingly, the expression of pluripotency-related genes was up-regulated in CPP-CARM1-treated mESCs. In addition, CPP-CARM1-treated mESC-derived embryoid bodies (EBs) showed an elevated expression of pluripotency-related genes and delayed spontaneous differentiation. This result suggests that the treatment of recombinant CPP-CARM1 protein elevates the expression of pluripotency-related genes of mESCs by epigenetic modification, and this protein-delivery system could be used to modify embryonic fate in reconstituted embryos with mESCs.
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University, Seoul 135-081, Korea.
authors.forename: Sara
authors.initials: S
authors.lastname: Choi
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University, Seoul 135-081, Korea.
authors.forename: Junghyun
authors.initials: J
authors.lastname: Jo
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University, Seoul 135-081, Korea.
authors.forename: Dong-Won
authors.initials: DW
authors.lastname: Seol
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University, Seoul 135-081, Korea ; Fertility Center of CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul 135-081, Korea.
authors.forename: Soo Kyung
authors.initials: SK
authors.lastname: Cha
authors.affiliation: CHA Stem Cell Institute, CHA University, Seoul 135-081, Korea.
authors.forename: Jeoung Eun
authors.initials: JE
authors.lastname: Lee
authors.affiliation: Department of Biomedical Science, College of Life Science, CHA University, Seoul 135-081, Korea ; Fertility Center of CHA Gangnam Medical Center, College of Medicine, CHA University, Seoul 135-081, Korea ; CHA Stem Cell Institute, CHA University, Seoul 135-081, Korea.
authors.forename: Dong Ryul
authors.initials: DR
authors.lastname: Lee
keywords: 3-germ layers; CARM1; Cell-penetrating peptide; Mouse embryonic stem cells; Pluripotent-related genes