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The use of whole-genome pooled shRNA libraries in loss-of-function screening in tissue culture models provides an effective means to identify novel factors acting in pathways of interest. Embryonic stem cells (ESCs) offer a unique opportunity to study processes involved in stem cell pluripotency and differentiation. Here, we report a genome-wide shRNA screen in ESCs to identify novel components involved in repression of the Gata6 locus, using a cell viability-based screen, which offers the benefits of stable shRNA integration and a robust and simple protocol for hit identification. Candidate factors identified were enriched for transcription factors and included known Polycomb proteins and other chromatin-modifying factors. We identified the protein Bcor, which is known to associate in complexes with the Polycomb protein Ring1B, and verified its importance in Gata6 repression in ESCs. Potential further applications of such a screening strategy could allow the identification of factors important for regulation of gene expression and pluripotency.

Original publication

DOI

10.1242/dev.094615

Type

Journal article

Journal

Development

Publication Date

10/2013

Volume

140

Pages

4110 - 4115

Keywords

Embryonic stem cells, Mouse, Polycomb, shRNA screen, Animals, Embryonic Stem Cells, GATA6 Transcription Factor, Mice, Mice, Knockout, RNA, Small Interfering