1. Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA

Correspondence to: Natalia Ivanova¹Ihor R. Lemischka¹ Correspondence and requests for materials should be addressed to N.I. (Email: nivanova@molbio.princeton.edu) or I.R.L. (Email: ilemischka@molbio.princeton.edu).

Abstract

We present an integrated approach to identify genetic mechanisms that control self-renewal in mouse embryonic stem cells. We use short hairpin RNA (shRNA) loss-of-function techniques to downregulate a set of gene products whose expression patterns suggest self-renewal regulatory functions. We focus on transcriptional regulators and identify seven genes for which shRNA-mediated depletion negatively affects self-renewal, including four genes with previously unrecognized roles in self-renewal. Perturbations of these gene products are combined with dynamic, global analyses of gene expression. Our studies suggest specific biological roles for these molecules and reveal the complexity of cell fate regulation in embryonic stem cells.

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