Open AccessDifferentiation of Mouse Embryonic Stem Cells after RNA Interference‐Mediated Silencing of OCT4 and Nanog
Author(s) -
Hough Shelley R.,
Clements Ian,
Welch Peter J.,
Wiederholt Kristin A.
Publication year - 2006
Publication title -
stem cells
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.159
H-Index - 229
eISSN - 1549-4918
pISSN - 1066-5099
DOI - 10.1634/stemcells.2005-0475
Subject(s) - homeobox protein nanog , biology , rna interference , rex1 , gene silencing , embryonic stem cell , microbiology and biotechnology , nanog homeobox protein , stem cell , cellular differentiation , induced pluripotent stem cell , genetics , gene , rna
RNA interference (RNAi) holds great promise as a tool to study the basic biology of stem cells or to direct differentiation in a specific manner. Barriers to achieving efficient and specific gene silencing in RNAi experiments include limitations in transfection efficiency and in the efficacy and specificity of RNAi silencing effectors. Here, we combine methods of efficient lipid‐mediated delivery with chemically modified RNAi compounds to silence genes related to pluripotency, in order to direct differentiation of mouse embryonic stem cells. After transfection of embryonic stem cells with OCT4‐ or Nanog‐targeted RNAi compounds, levels of OCT4 or Nanog transcript and protein were reduced accordingly. Reduction in OCT4 expression correlated with induction of trophectoderm genes Cdx2 , Hand1 , and PL‐1 , with formation of cells with trophoblast giant cell phenotype after 6 days. Reduction in Nanog expression correlated with induction of extraembryonic endoderm genes GATA4 , GATA6 , and laminin B1 , with subsequent generation of groups of cells with parietal endoderm phenotype. Our results indicate that transient inhibition of OCT4 or Nanog by RNAi compounds is sufficient to induce differentiation toward extraembryonic lineages, which supports the model that these transcription factors function in a dose‐dependent manner to influence cell fate.