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Stem Cells. 2005 Mar;23(3):299-305.

High-efficiency RNA interference in human embryonic stem cells.

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Harvard Stem Cell Institute and Harvard Medical School, Department of Biological Chemistry and Molecular Pharmacology and Division of Hematology/Oncology, Children's Hospital, Boston, Massachusetts 02115, USA.


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.

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