title: Zfp296 is a novel, pluripotent-specific reprogramming factor.
abstract: Expression of the four transcription factors Oct4, Sox2, Klf4, and c-Myc (OSKM) is sufficient to reprogram somatic cells into induced pluripotent stem (iPSCs). However, this process is slow and inefficient compared with the fusion of somatic cells with embryonic stem cells (ESCs), indicating that ESCs express additional factors that can enhance the efficiency of reprogramming. We had previously developed a method to detect and isolate early neural induction intermediates during the differentiation of mouse ESCs. Using the gene expression profiles of these intermediates, we identified 23 ESC-specific transcripts and tested each for the ability to enhance iPSC formation. Of the tested factors, zinc finger protein 296 (Zfp296) led to the largest increase in mouse iPSC formation. We confirmed that Zfp296 was specifically expressed in pluripotent stem cells and germ cells. Zfp296 in combination with OSKM induced iPSC formation earlier and more efficiently than OSKM alone. Through mouse chimera and teratoma formation, we demonstrated that the resultant iPSCs were pluripotent. We showed that Zfp296 activates transcription of the Oct4 gene via the germ cell-specific conserved region 4 (CR4), and when overexpressed in mouse ESCs leads to upregulation of Nanog expression and downregulation of the expression of differentiation markers, including Sox17, Eomes, and T, which is consistent with the observation that Zfp296 enhances the efficiency of reprogramming. In contrast, knockdown of Zfp296 in ESCs leads to the expression of differentiation markers. Finally, we demonstrated that expression of Zfp296 in ESCs inhibits, but does not block, differentiation into neural cells.
authors.affiliation: Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine, Münster, Germany.
authors.forename: Gerrit
authors.initials: G
authors.lastname: Fischedick
authors.forename: Diana C
authors.initials: DC
authors.lastname: Klein
authors.forename: Guangming
authors.initials: G
authors.lastname: Wu
authors.forename: Daniel
authors.initials: D
authors.lastname: Esch
authors.forename: Susanne
authors.initials: S
authors.lastname: Höing
authors.forename: Dong Wook
authors.initials: DW
authors.lastname: Han
authors.forename: Peter
authors.initials: P
authors.lastname: Reinhardt
authors.forename: Kerstin
authors.initials: K
authors.lastname: Hergarten
authors.forename: Natalia
authors.initials: N
authors.lastname: Tapia
authors.forename: Hans R
authors.initials: HR
authors.lastname: Schöler
authors.forename: Jared L
authors.initials: JL
authors.lastname: Sterneckert
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000378:metabolism; D054337:Cell Dedifferentiation; D002454:Cell Differentiation; D002678:Chimera; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D053595:Embryonic Stem Cells / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D057809:HEK293 Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000502:physiology* / Q000637:transplantation; D000090062:Kruppel-Like Factor 4; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D016513:Mice, SCID; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D009928:Organ Specificity; D011401:Promoter Regions, Genetic; D013724:Teratoma / Q000473:pathology; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology
chemical_list: D000943:Antigens, Differentiation; D004268:DNA-Binding Proteins; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D014157:Transcription Factors; C421709:zinc finger protein 296, mouse