Stem Cells
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First published online February 22, 2007
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2006-0825v1
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Submitted on December 28, 2006
Accepted on February 12, 2007

Embryonic Stem Cells

Conditional gene expression in human embryonic stem cells

Ludovic Vallier 1*, Morgan Alexander 1, Roger Pedersen 1

1 Department of Surgery and Cambridge Institute for Medical Research, Addenbrooke's Hospital, University of Cambridge, Cambridge, United Kingdom

* To whom correspondence should be addressed. E-mail: lv225{at}cam.ac.uk.


  Abstract

Human Embryonic Stem Cells (hESCs) possess unique properties for studying mechanisms controlling cell fate commitment during early mammalian development. Gain of function is a common strategy to study the function of specific genes involved in these mechanisms. However, transgene toxicity can be a major limitation, especially with factors influencing proliferation or differentiation. Here, we describe an efficient method based on the inducible recombinase Cre-ERT2 for conditional gene expression in hESCs and their differentiated derivatives. Using this approach, we have established several hESC sub-lines inducible for the expression of the enhanced Green Fluorescent Protein and the TGF{beta} growth factor family member Nodal. Together these results demonstrate that Cre-ERT2 can be used to control gene expression in undifferentiated and differentiated cells, thereby providing the first conditional transgene expression system that works effectively in hESCs.

Key Words. Human embryonic stem cells, Cre recombinase, Tamoxifen, Nodal, inducible expression, pluripotency




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