High-efficiency siRNA-based gene knockdown in human embryonic stem cells

• Published in: RNA (Cambridge) Vol. 16; no. 12; pp. 2564 - 2569
• Main Authors: Ma, Yinghong, Jin, Jianyu, Dong, Chunsheng, Cheng, Ee-Chun, Lin, Haifan, Huang, Yingqun, Qiu, Caihong
• Format: Journal Article
• Language: English
• Published: United States Cold Spring Harbor Laboratory Press 01.12.2010
• Subjects: Cell Count; Cell Differentiation - drug effects; Cell Proliferation - drug effects; Cell Survival - drug effects; Cells, Cultured; Efficiency; Embryonic Stem Cells - cytology; Embryonic Stem Cells - drug effects; Embryonic Stem Cells - metabolism; Embryonic Stem Cells - physiology; Gene Knockdown Techniques - methods; Genes, Reporter; Green Fluorescent Proteins - genetics; Green Fluorescent Proteins - metabolism; Humans; Method; Octamer Transcription Factor-3 - antagonists & inhibitors; Octamer Transcription Factor-3 - genetics; Octamer Transcription Factor-3 - metabolism; RNA, Small Interfering - genetics; RNA, Small Interfering - metabolism; RNA, Small Interfering - pharmacology; Transfection - methods
• ISSN: 1355-8382; 1469-9001
• DOI: 10.1261/rna.2350710

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.