title: Two miRNA clusters reveal alternative paths in late-stage reprogramming.
abstract: Ectopic expression of specific factors such as Oct4, Sox2, and Klf4 (OSK) is sufficient to reprogram somatic cells into induced pluripotent stem cells (iPSCs). In this study, we examine the paths taken by cells during the reprogramming process by following the transcriptional activation of two pluripotent miRNA clusters (mir-290 and mir-302) in individual cells in vivo and in vitro with knockin reporters. During embryonic development and embryonic stem cell differentiation, all cells sequentially expressed mir-290 and mir-302. In contrast, during OSK-induced reprogramming, cells activated the miRNA loci in a stochastic, nonordered manner. However, the addition of Sall4 to the OSK cocktail led to a consistent reverse sequence of locus activation (mir-302 then mir-290) and increased reprogramming efficiency. These results demonstrate that cells can follow multiple paths during the late stages of reprogramming, and that the trajectory of any individual cell is strongly influenced by the combination of factors introduced.
authors.affiliation: The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Center for Reproductive Sciences, University of California, San Francisco, San Francisco, California, 94143, USA; Department of Urology, University of California, San Francisco, San Francisco, California, 94143, USA.
authors.forename: Ronald J
authors.initials: RJ
authors.lastname: Parchem
authors.affiliation: The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Center for Reproductive Sciences, University of California, San Francisco, San Francisco, California, 94143, USA; Department of Urology, University of California, San Francisco, San Francisco, California, 94143, USA.
authors.forename: Julia
authors.initials: J
authors.lastname: Ye
authors.affiliation: The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Center for Reproductive Sciences, University of California, San Francisco, San Francisco, California, 94143, USA; Department of Urology, University of California, San Francisco, San Francisco, California, 94143, USA.
authors.forename: Robert L
authors.initials: RL
authors.lastname: Judson
authors.affiliation: The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Center for Reproductive Sciences, University of California, San Francisco, San Francisco, California, 94143, USA; Department of Urology, University of California, San Francisco, San Francisco, California, 94143, USA.
authors.forename: Marie F
authors.initials: MF
authors.lastname: LaRussa
authors.affiliation: The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Center for Reproductive Sciences, University of California, San Francisco, San Francisco, California, 94143, USA; Department of Urology, University of California, San Francisco, San Francisco, California, 94143, USA.
authors.forename: Raga
authors.initials: R
authors.lastname: Krishnakumar
authors.affiliation: The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Center for Reproductive Sciences, University of California, San Francisco, San Francisco, California, 94143, USA.
authors.forename: Amy
authors.initials: A
authors.lastname: Blelloch
authors.affiliation: The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Center for Reproductive Sciences, University of California, San Francisco, San Francisco, California, 94143, USA; Department of Neurology, University of California, San Francisco, San Francisco, California, 94143, USA.
authors.forename: Michael C
authors.initials: MC
authors.lastname: Oldham
authors.affiliation: The Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, Center for Reproductive Sciences, University of California, San Francisco, San Francisco, California, 94143, USA; Department of Urology, University of California, San Francisco, San Francisco, California, 94143, USA. Electronic address: blellochr@stemcell.ucsf.edu.
authors.forename: Robert
authors.initials: R
authors.lastname: Blelloch
mesh_terms: D000818:Animals; D065150:Cellular Reprogramming / Q000235:genetics / Q000502:physiology*; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D005260:Female; D000090062:Kruppel-Like Factor 4; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism
chemical_list: D004268:DNA-Binding Proteins; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; C547384:MIRN290 microRNA, mouse; C558265:MIRN302 microRNA, mouse; D035683:MicroRNAs; C509167:Sall4 protein, mouse; D014157:Transcription Factors