High-efficiency RNA interference in human embryonic stem cells.
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.
Zaehres
Holm
H
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.
Lensch
M William
MW
Daheron
Laurence
L
Stewart
Sheila A
SA
Itskovitz-Eldor
Joseph
J
Daley
George Q
GQ
D000954:Antigens, Surface; D002454:Cell Differentiation / Q000502:physiology; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000235:genetics / Q000502:physiology; D004622:Embryo, Mammalian / Q000166:cytology*; D015870:Gene Expression / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D005822:Genetic Vectors / Q000235:genetics; D015292:Glycoprotein Hormones, alpha Subunit / Q000235:genetics; D006023:Glycoproteins / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics; D018398:Homeodomain Proteins / Q000235:genetics / Q000502:physiology; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; D039904:Pluripotent Stem Cells / Q000378:metabolism / Q000502:physiology*; D011509:Proteoglycans; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics; D012190:Retroviridae / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012709:Serum Albumin / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology*; D014162:Transfection
D000954:Antigens, Surface; D004268:DNA-Binding Proteins; D015292:Glycoprotein Hormones, alpha Subunit; D006023:Glycoproteins; D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D011509:Proteoglycans; D034741:RNA, Small Interfering; D012709:Serum Albumin; C107064:TRA-1-60 antigen, human; D014157:Transcription Factors; D049452:Green Fluorescent Proteins