High‐Efficiency RNA Interference in Human Embryonic Stem Cells

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.