title: Specific knockdown of Oct4 and beta2-microglobulin expression by RNA interference in human embryonic stem cells and embryonic carcinoma cells.
abstract: We have used RNA interference (RNAi) to downregulate beta2-microglobulin and Oct4 in human embryonal carcinoma (hEC) cells and embryonic stem (hES) cells, demonstrating that RNAi is an effective tool for regulating specific gene activity in these human stem cells. The knockdown of Oct4 but not beta2-microglobulin expression in both EC and ES cells resulted in their differentiation, as indicated by a marked change in morphology, growth rate, and surface antigen phenotype, with respect to SSEA1, SSEA3, and TRA-1-60 expression. Expression of hCG and Gcm1 was also induced following knockdown of Oct4 expression, in both 2102Ep hEC cells and in H7 and H14 hES cells, consistent with the conclusion that, as in the mouse, Oct4 is required to maintain the undifferentiated stem cell state, and that differentiation to trophectoderm occurs in its absence. NTERA2 hEC cells also differentiated, but not to trophectoderm, suggesting their equivalence to a later stage of embryogenesis than other hEC and hES cells.
authors.affiliation: The Centre for Stem Cell Biology, University of Sheffield, Western Bank, S10 2TN, U.K.
authors.forename: Maryam M
authors.initials: MM
authors.lastname: Matin
authors.forename: James R
authors.initials: JR
authors.lastname: Walsh
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Gokhale
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Draper
authors.forename: Ahmad R
authors.initials: AR
authors.lastname: Bahrami
authors.forename: Ian
authors.initials: I
authors.lastname: Morton
authors.forename: Harry D
authors.initials: HD
authors.lastname: Moore
authors.forename: Peter W
authors.initials: PW
authors.lastname: Andrews
mesh_terms: D000954:Antigens, Surface / Q000235:genetics; D002454:Cell Differentiation / Q000235:genetics; D002460:Cell Line; D049109:Cell Proliferation; D006063:Chorionic Gonadotropin / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics*; D015536:Down-Regulation / Q000235:genetics*; D004475:Ectoderm / Q000378:metabolism; D054278:Embryonal Carcinoma Stem Cells; D047108:Embryonic Development / Q000235:genetics; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D006801:Humans; D014411:Neoplastic Stem Cells / Q000378:metabolism*; D009479:Neuropeptides / Q000235:genetics; D009687:Nuclear Proteins; D050814:Octamer Transcription Factor-3; D039904:Pluripotent Stem Cells / Q000378:metabolism*; D034622:RNA Interference / Q000502:physiology*; D014157:Transcription Factors / Q000235:genetics*; D001613:beta 2-Microglobulin / Q000235:genetics*
chemical_list: D000954:Antigens, Surface; D006063:Chorionic Gonadotropin; D004268:DNA-Binding Proteins; C491089:GCM1 protein, human; D009479:Neuropeptides; D009687:Nuclear Proteins; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D014157:Transcription Factors; D001613:beta 2-Microglobulin