Members of the miR-290 cluster modulate in vitro differentiation of mouse embryonic stem cells

Differentiation. 2009 Sep-Oct;78(2-3):69-78. doi: 10.1016/j.diff.2009.06.003. Epub 2009 Jul 22.

Abstract

We report the biological effects of miR-290 cluster via gain-of-function or loss-of-function experiments in mouse embryonic stem cells (ESCs) cultured under differentiation conditions. Under these conditions we found that overexpression of miR-290 cluster in ESCs cannot prevent downregulation of Oct-4, but inhibition results in earlier downregulation of Oct-4 compared with the negative control. In consistence with previous findings that report ectopic expression of Brachyury during gastrulation in Argonaute-2 KO mice due to impaired miRNA function, we show that miR-290 cluster regulates negatively differentiation of ESCs towards mesodermal and germ cell lineage. These results suggest that although incapable to maintain pluripotent state alone, miR-290 cluster inhibits ESC differentiation and it is involved in the pathways controlling mesoderm and primordial germ cell differentiation. Finally, we provide proofs that members of this cluster target Dkk-1 gene, a Wnt pathway inhibitor, and affect this pathway, which can partially explain why miR-290 cluster favours pluripotency against differentiation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation / physiology*
  • Cells, Cultured
  • Chromosomal Proteins, Non-Histone
  • Embryonic Stem Cells / cytology*
  • Gene Expression Profiling
  • Germ Cells / physiology*
  • Intercellular Signaling Peptides and Proteins / genetics
  • Intercellular Signaling Peptides and Proteins / metabolism*
  • Luciferases / metabolism
  • Mesoderm / cytology
  • Mesoderm / physiology
  • Mice
  • MicroRNAs / physiology*
  • Octamer Transcription Factor-3 / genetics
  • Octamer Transcription Factor-3 / metabolism
  • Pluripotent Stem Cells
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism*
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Chromosomal Proteins, Non-Histone
  • Dkk1 protein, mouse
  • Dppa3 protein, mouse
  • Intercellular Signaling Peptides and Proteins
  • MicroRNAs
  • Octamer Transcription Factor-3
  • Pou5f1 protein, mouse
  • RNA, Messenger
  • Repressor Proteins
  • Luciferases