Abstract
RNA interference methodology suppresses gene expression, thus mimicking loss-of-function mutation and enabling in vitro and in vivo gene function analysis. In this study, we used retroviral and lentiviral vectors to deliver small interfering RNAs and report high-efficiency silencing of a green fluorescent protein (GFP) trans gene and the stem cell-specific transcription factors Oct4/POU5F1 and Nanog in human embryonic stem cells. Gene knockdown of Oct4 and Nanog promotes differentiation, thereby demonstrating a role for these factors in human embryonic stem cell self-renewal.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
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Research Support, U.S. Gov't, Non-P.H.S.
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Research Support, U.S. Gov't, P.H.S.
MeSH terms
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Antigens, Surface
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Cell Differentiation / physiology
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Cell Line
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / physiology
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Embryo, Mammalian / cytology*
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Gene Expression / genetics
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Gene Expression Regulation, Developmental
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Genetic Vectors / genetics
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Glycoprotein Hormones, alpha Subunit / genetics
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Glycoproteins / metabolism
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Green Fluorescent Proteins / genetics
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Homeodomain Proteins / genetics
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Homeodomain Proteins / physiology
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Humans
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Lentivirus / genetics
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Nanog Homeobox Protein
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Octamer Transcription Factor-3
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Pluripotent Stem Cells / metabolism
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Pluripotent Stem Cells / physiology*
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Proteoglycans
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RNA Interference*
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RNA, Small Interfering / genetics
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Retroviridae / genetics
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Reverse Transcriptase Polymerase Chain Reaction
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Serum Albumin / genetics
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Transcription Factors / genetics
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Transcription Factors / physiology*
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Transfection
Substances
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Antigens, Surface
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DNA-Binding Proteins
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Glycoprotein Hormones, alpha Subunit
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Glycoproteins
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Homeodomain Proteins
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NANOG protein, human
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Nanog Homeobox Protein
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Octamer Transcription Factor-3
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POU5F1 protein, human
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Proteoglycans
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RNA, Small Interfering
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Serum Albumin
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TRA-1-60 antigen, human
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Transcription Factors
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Green Fluorescent Proteins