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 tra...

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Bibliographic Details
Published in:Genesis (New York, N.Y. : 2000) N.Y. : 2000), 2003-09, Vol.37 (1), p.18-24
Main Authors: Velkey, J. Matthew, O'Shea, K. Sue
Format: Article
Language:English
Subjects:
Animals
Cell Differentiation
Cell Lineage
Cell Separation
Cells, Cultured
differentiation
DNA-Binding Proteins - genetics
Ectoderm - cytology
Embryo, Mammalian - cytology
embryonic stem cells
Flow Cytometry
Genes, Reporter
Green Fluorescent Proteins
Immunohistochemistry
Luminescent Proteins - metabolism
Mice
Models, Genetic
Oct-4 protein
Oct4
Octamer Transcription Factor-3
Oligonucleotides, Antisense - chemistry
Plasmids - metabolism
Promoter Regions, Genetic
Reverse Transcriptase Polymerase Chain Reaction
RNA Interference
RNA Polymerase III - genetics
Stem Cells - cytology
Stem Cells - metabolism
Time Factors
Transcription Factors
Transfection
trophoblast
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Summary: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. genesis 37:18–24, 2003. © 2003 Wiley‐Liss, Inc.
ISSN:1526-954X
1526-968X
DOI:10.1002/gene.10218