Dppa3 expression is critical for generation of fully reprogrammed iPS cells and maintenance of Dlk1-Dio3 imprinting

Nat Commun. 2015 Jan 23:6:6008. doi: 10.1038/ncomms7008.

Abstract

Reprogramming of mouse somatic cells into induced pluripotent stem cells (iPSCs) often generates partially reprogrammed iPSCs (pre-iPSCs), low-grade chimera forming iPSCs (lg-iPSCs) and fully reprogrammed, high-grade chimera production competent iPSCs (hg-iPSCs). Lg-iPSC transcriptome analysis revealed misregulated Dlk1-Dio3 cluster gene expression and subsequently the imprinting defect at the Dlk1-Dio3 locus. Here, we show that germ-cell marker Dppa3 is present only in lg-iPSCs and hg-iPSCs, and that induction with exogenous Dppa3 enhances reprogramming kinetics, generating all hg-iPSCs, similar to vitamin C (Vc). Conversely, Dppa3-null fibroblasts show reprogramming block at pre-iPSCs state and Dlk1-Dio3 imprinting defect. At the molecular level, we show that Dppa3 is associated with Dlk1-Dio3 locus and identify that Dppa3 maintains imprinting by antagonizing Dnmt3a binding. Our results further show molecular parallels between Dppa3 and Vc in Dlk1-Dio3 imprinting maintenance and suggest that early activation of Dppa3 is one of the cascades through which Vc facilitates the generation of fully reprogrammed iPSCs.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Ascorbic Acid / metabolism
  • Calcium-Binding Proteins
  • Chromosomal Proteins, Non-Histone
  • Crosses, Genetic
  • DNA Methylation
  • Female
  • Fibroblasts / metabolism
  • Gene Expression Profiling
  • Gene Expression Regulation*
  • Genomic Imprinting
  • Germ Cells / cytology
  • Green Fluorescent Proteins / metabolism
  • Induced Pluripotent Stem Cells / metabolism*
  • Intercellular Signaling Peptides and Proteins / metabolism*
  • Iodide Peroxidase / metabolism*
  • Kinetics
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Mice, Transgenic
  • Protein Binding
  • Repressor Proteins / metabolism*
  • Retroviridae / metabolism

Substances

  • Calcium-Binding Proteins
  • Chromosomal Proteins, Non-Histone
  • Dlk1 protein, mouse
  • Dppa3 protein, mouse
  • Intercellular Signaling Peptides and Proteins
  • Repressor Proteins
  • Green Fluorescent Proteins
  • iodothyronine deiodinase type III
  • Iodide Peroxidase
  • Ascorbic Acid