Abstract
Loss-of-function studies in human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) via nonviral approaches have been largely unsuccessful. Here we report a simple and cost-effective method for high-efficiency delivery of plasmids and siRNAs into hESCs and iPSCs. Using this method for siRNA delivery, we achieve >90% reduction in the expression of the stem cell factors Oct4 and Lin28, and observe cell morphological and staining pattern changes, characteristics of hESC differentiation, as a result of Oct4 knockdown.
Publication types
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Evaluation Study
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Research Support, Non-U.S. Gov't
MeSH terms
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Cell Count
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Cell Differentiation / drug effects
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Cell Proliferation / drug effects
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Cell Survival / drug effects
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Cells, Cultured
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Efficiency
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Embryonic Stem Cells / cytology
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Embryonic Stem Cells / drug effects*
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Embryonic Stem Cells / metabolism*
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Embryonic Stem Cells / physiology
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Gene Knockdown Techniques / methods*
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Genes, Reporter
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Green Fluorescent Proteins / genetics
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Green Fluorescent Proteins / metabolism
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Humans
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Octamer Transcription Factor-3 / antagonists & inhibitors
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Octamer Transcription Factor-3 / genetics
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Octamer Transcription Factor-3 / metabolism
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RNA, Small Interfering / genetics
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RNA, Small Interfering / metabolism
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RNA, Small Interfering / pharmacology*
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Transfection / methods
Substances
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Octamer Transcription Factor-3
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POU5F1 protein, human
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RNA, Small Interfering
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Green Fluorescent Proteins