title: Deterministic direct reprogramming of somatic cells to pluripotency.
abstract: Somatic cells can be inefficiently and stochastically reprogrammed into induced pluripotent stem (iPS) cells by exogenous expression of Oct4 (also called Pou5f1), Sox2, Klf4 and Myc (hereafter referred to as OSKM). The nature of the predominant rate-limiting barrier(s) preventing the majority of cells to successfully and synchronously reprogram remains to be defined. Here we show that depleting Mbd3, a core member of the Mbd3/NuRD (nucleosome remodelling and deacetylation) repressor complex, together with OSKM transduction and reprogramming in naive pluripotency promoting conditions, result in deterministic and synchronized iPS cell reprogramming (near 100% efficiency within seven days from mouse and human cells). Our findings uncover a dichotomous molecular function for the reprogramming factors, serving to reactivate endogenous pluripotency networks while simultaneously directly recruiting the Mbd3/NuRD repressor complex that potently restrains the reactivation of OSKM downstream target genes. Subsequently, the latter interactions, which are largely depleted during early pre-implantation development in vivo, lead to a stochastic and protracted reprogramming trajectory towards pluripotency in vitro. The deterministic reprogramming approach devised here offers a novel platform for the dissection of molecular dynamics leading to establishing pluripotency at unprecedented flexibility and resolution.
authors.affiliation: The Department of Molecular Genetics, Weizmann Institute of Science, Rehovot 76100, Israel.
authors.forename: Yoach
authors.initials: Y
authors.lastname: Rais
authors.forename: Asaf
authors.initials: A
authors.lastname: Zviran
authors.forename: Shay
authors.initials: S
authors.lastname: Geula
authors.forename: Ohad
authors.initials: O
authors.lastname: Gafni
authors.forename: Elad
authors.initials: E
authors.lastname: Chomsky
authors.forename: Sergey
authors.initials: S
authors.lastname: Viukov
authors.forename: Abed AlFatah
authors.initials: AA
authors.lastname: Mansour
authors.forename: Inbal
authors.initials: I
authors.lastname: Caspi
authors.forename: Vladislav
authors.initials: V
authors.lastname: Krupalnik
authors.forename: Mirie
authors.initials: M
authors.lastname: Zerbib
authors.forename: Itay
authors.initials: I
authors.lastname: Maza
authors.forename: Nofar
authors.initials: N
authors.lastname: Mor
authors.forename: Dror
authors.initials: D
authors.lastname: Baran
authors.forename: Leehee
authors.initials: L
authors.lastname: Weinberger
authors.forename: Diego A
authors.initials: DA
authors.lastname: Jaitin
authors.forename: David
authors.initials: D
authors.lastname: Lara-Astiaso
authors.forename: Ronnie
authors.initials: R
authors.lastname: Blecher-Gonen
authors.forename: Zohar
authors.initials: Z
authors.lastname: Shipony
authors.forename: Zohar
authors.initials: Z
authors.lastname: Mukamel
authors.forename: Tzachi
authors.initials: T
authors.lastname: Hagai
authors.forename: Shlomit
authors.initials: S
authors.lastname: Gilad
authors.forename: Daniela
authors.initials: D
authors.lastname: Amann-Zalcenstein
authors.forename: Amos
authors.initials: A
authors.lastname: Tanay
authors.forename: Ido
authors.initials: I
authors.lastname: Amit
authors.forename: Noa
authors.initials: N
authors.lastname: Novershtern
authors.forename: Jacob H
authors.initials: JH
authors.lastname: Hanna
mesh_terms: D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000235:genetics / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000235:genetics; D053595:Embryonic Stem Cells; D005260:Female; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000502:physiology*; D000090062:Kruppel-Like Factor 4; D008297:Male; D051379:Mice; D008954:Models, Biological; D014157:Transcription Factors / Q000235:genetics
chemical_list: D004268:DNA-Binding Proteins; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; C492013:Mbd3 protein, mouse; D014157:Transcription Factors
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