High-Efficiency RNA Interference in Human Embryonic Stem Cells

Authors

  • Holm Zaehres,

    1. 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, USA
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  • M. William Lensch,

    1. 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, USA
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  • Laurence Daheron,

    1. 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, USA
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  • Sheila A. Stewart,

    1. Washington University School of Medicine, St. Louis, Missouri, USA
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  • Joseph Itskovitz-Eldor,

    1. Technion, Rambam Medical Center, Haifa, Israel
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  • George Q. Daley M.D., Ph.D.

    Corresponding author
    1. 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, USA
    • 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. Telephone: 617-919-2013; Fax: 617-730-0222
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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.

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