title: CHIR99021 enhances Klf4 Expression through β-Catenin Signaling and miR-7a Regulation in J1 Mouse Embryonic Stem Cells.
abstract: Understanding the mechanisms that regulate pluripotency of embryonic stem cells (ESCs) is important to ensure their safe clinical use. CHIR99021 (CHIR)-induced activation of Wnt/β-catenin signaling promotes self-renewal in mouse ESCs (mESCs). β-catenin functions individually or cooperates with transcription factors to activate stemness factors such as c-Myc, Esrrb, Pou5f1, and Nanog. However the relationship between the core pluripotent factor, Kruppel-like factor 4 (also known as GKLF or EZF) and Wnt/β-catenin signaling, remains ambiguous in J1 mESCs. DNA microarray analysis revealed that CHIR-treatment promoted pluripotency-maintaining transcription factors and repressed germ layer specification markers. CHIR also promoted genes related to the development of extracellular regions and the plasma membrane to maintain pluripotency of J1 mESCs. Among the CHIR-regulated genes, Klf4 has not been reported previously. We identified a novel cis element in the Klf4 gene that was activated by β-catenin in J1 mESCs. We determined that β-catenin interacted with this cis element, identifying Klf4 as a β-catenin target gene in this context. Moreover, several microRNAs that targeted the 3'-UTR of Klf4 mRNA were identified, with miR-7a being down-regulated by CHIR in a β-catenin-independent manner in J1 mESCs. These data collectively suggest that CHIR enhances Klf4 expression by repressing miR-7a expression or canonical Wnt pathway activation.
authors.affiliation: College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, China.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A& F University, Yangling, Shaanxi, China.
authors.forename: Zhiying
authors.initials: Z
authors.lastname: Ai
authors.affiliation: College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, China.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A& F University, Yangling, Shaanxi, China.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Shao
authors.affiliation: College of Veterinary Medicine, Northwest A& F University, Yangling, Shaanxi, China.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A& F University, Yangling, Shaanxi, China.
authors.forename: Yongyan
authors.initials: Y
authors.lastname: Wu
authors.affiliation: College of Veterinary Medicine, Northwest A& F University, Yangling, Shaanxi, China.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A& F University, Yangling, Shaanxi, China.
authors.forename: Mengying
authors.initials: M
authors.lastname: Yu
authors.affiliation: College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, China.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A& F University, Yangling, Shaanxi, China.
authors.forename: Juan
authors.initials: J
authors.lastname: Du
authors.affiliation: College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, China.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A& F University, Yangling, Shaanxi, China.
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Shi
authors.affiliation: College of Life Sciences, Northwest A&F University, Yangling, Shaanxi, China.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A& F University, Yangling, Shaanxi, China.
authors.forename: Xinglong
authors.initials: X
authors.lastname: Shi
authors.affiliation: College of Veterinary Medicine, Northwest A& F University, Yangling, Shaanxi, China.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A& F University, Yangling, Shaanxi, China.
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: College of Veterinary Medicine, Northwest A& F University, Yangling, Shaanxi, China.|Key Laboratory of Animal Biotechnology, Ministry of Agriculture, Northwest A& F University, Yangling, Shaanxi, China.
authors.forename: Zekun
authors.initials: Z
authors.lastname: Guo
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D005786:Gene Expression Regulation; D063990:Gene Ontology; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors / Q000819:agonists / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D058977:Molecular Sequence Annotation; D008969:Molecular Sequence Data; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics / Q000378:metabolism; D011725:Pyridines / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology*; D011960:Receptors, Estrogen / Q000235:genetics / Q000378:metabolism; D060449:Wnt Signaling Pathway / Q000187:drug effects; D051176:beta Catenin / Q000235:genetics* / Q000378:metabolism
chemical_list: D020413:3' Untranslated Regions; C495267:CTNNB1 protein, mouse; C473711:Chir 99021; C098925:Esrrb protein, mouse; D018398:Homeodomain Proteins; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D051741:Kruppel-Like Transcription Factors; C549243:MIRN7 microRNA, mouse; D035683:MicroRNAs; C489428:Myc protein, mouse; D000071317:Nanog Homeobox Protein; C475540:Nanog protein, mouse; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D047428:Protein Kinase Inhibitors; D016271:Proto-Oncogene Proteins c-myc; D011725:Pyridines; D011743:Pyrimidines; D011960:Receptors, Estrogen; D051176:beta Catenin