Oct4 RNA interference induces trophectoderm differentiation in mouse embryonic stem cells.
We examined whether suppression of Oct4 via RNA interference (RNAi) would affect embryonic stem (ES) cell lineage choice. Cells were transfected with plasmids containing an independently expressed reporter gene and an RNA polymerase type III promoter to constitutively express small stem-loop RNA transcripts corresponding to Oct4 mRNA. Cells transfected with Oct4 RNAi constructs demonstrated reduced levels of Oct4 mRNA and exhibited characteristics of trophectodermal differentiation. These findings support the critical role of Oct4 in regulating stem cell identity and suggest that future experiments using RNAi in ES cells can elucidate the roles of other genes affecting lineage specification during differentiation.
D000818:Animals; D002454:Cell Differentiation; D019070:Cell Lineage; D002469:Cell Separation; D002478:Cells, Cultured; D004268:DNA-Binding Proteins / Q000235:genetics*; D004475:Ectoderm / Q000166:cytology*; D004622:Embryo, Mammalian / Q000166:cytology; D005434:Flow Cytometry; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D007150:Immunohistochemistry; D008164:Luminescent Proteins / Q000378:metabolism; D051379:Mice; D008957:Models, Genetic; D050814:Octamer Transcription Factor-3; D016376:Oligonucleotides, Antisense / Q000737:chemistry; D010957:Plasmids / Q000378:metabolism; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D012320:RNA Polymerase III / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism; D013997:Time Factors; D014157:Transcription Factors; D014162:Transfection
D004268:DNA-Binding Proteins; D008164:Luminescent Proteins; D050814:Octamer Transcription Factor-3; D016376:Oligonucleotides, Antisense; C494174:Pou5f1 protein, mouse; D014157:Transcription Factors; D049452:Green Fluorescent Proteins; D012320:RNA Polymerase III