In Vitro- and In Vivo-Induced Transgene Expression in Human Embryonic Stem Cells and Derivatives

Abstract

The use of human embryonic stem cells (hESCs) as a research and therapeutic tool will be facilitated by conditional gene expression. Here, we report drug-induced transgene expression, both in vitro and in vivo, from a tet-on hESC line with >95% purity. Using green fluorescent protein as an indicator, we demonstrated that the tet-on system allowed a tight control of the gene expression in both undifferentiated hESCs and differentiated cells of the three germ layers. More importantly, after the cells were transplanted into animals, the gene expression remained to be regulated by an orally administered drug. These results provide a technical basis for regulation of gene expression in hESCs and derivatives in vitro and in vivo. STEM CELLS 2008;26:525-533 Disclosure of potential conflicts of interest is found at the end of this article.

A Simple and Efficient System for Regulating Gene Expression in Human Pluripotent Stem Cells and Derivatives

“…Efforts have been made to produce transgenic hPSC lines with regulatable gene expression (Du et al, 2009; Vigna et al, 2002; Xia et al, 2008; Zhou et al, 2007). While expression of transgenes (often GFP) can be regulated in stem cells and their differentiated progenies at an early stage, more than often transgenes are no longer expressed and regulatable in mature cells, including neurons (Cherry et al, 2000; Ellis, 2005; Laker et al, 1998; Yao et al, 2004).…”

Differential Effects of Hepatocyte Nuclear Factor 4α Isoforms on Tumor Growth and T-Cell Factor 4/AP-1 Interactions in Human Colorectal Cancer Cells

“…The expression vector containing the reverse tetracycline transcriptional activator (rtTA) pCAG-rtTA (pCAG-rtTA-IR-PURO) and the Tetresponsive red fluorescent protein (RFP) reporter construct pTRE-RFP were gifts from Chee-Gee Liew (48,49).…”

MSX2 mediates entry of human pluripotent stem cells into mesendoderm by simultaneously suppressing SOX2 and activating NODAL signaling

“…To explore the function of MSX2 in fate determination of hPSCs, we overexpressed MSX2 in hESCs using a previously described doxcycline (DOX) inducible lentiviral expression system and assessed its effect [37]. We used a GFP-MSX2 fusion gene which allowed us to monitor its expression in hESCs in real time (Supplementary information, Figure S1A).…”