Members of the miR-290 cluster modulate in vitro differentiation of mouse embryonic stem cells.
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.
Zovoilis
Athanasios
A
Institute of Human Genetics, University of Goettingen, Heinrich-Dueker-Weg 12, D-37073 Goettingen, Germany. azovoil@gwdg.de
Smorag
Lukasz
L
Pantazi
Angeliki
A
Engel
Wolfgang
W
D000818:Animals; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D002868:Chromosomal Proteins, Non-Histone; D053595:Embryonic Stem Cells / Q000166:cytology*; D020869:Gene Expression Profiling; D005854:Germ Cells / Q000502:physiology*; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D008156:Luciferases / Q000378:metabolism; D008648:Mesoderm / Q000166:cytology / Q000502:physiology; D051379:Mice; D035683:MicroRNAs / Q000502:physiology*; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D039904:Pluripotent Stem Cells; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D002868:Chromosomal Proteins, Non-Histone; C501127:Dkk1 protein, mouse; C462180:Dppa3 protein, mouse; D036341:Intercellular Signaling Peptides and Proteins; D035683:MicroRNAs; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D012333:RNA, Messenger; D012097:Repressor Proteins; D008156:Luciferases