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  current news   Press   selected story    
     
  24th November 2010  
 

A genome-wide RNAi screen reveals determinants of human ES cell identity

 
 




Authors
Na-Yu Chia1,2, , Yun-Shen Chan1,3,*, Bo Feng1,*, Xinyi Lu1,4, Yuriy L. Orlov5, Dimitri Moreau6, Pankaj Kumar6, Lin Yang1, Jianming Jiang1, Mei-Sheng Lau1, Mikael Huss5, Boon-Seng Soh7, Petra Kraus7, Pin Li7, Thomas Lufkin7, Bing Lim7,8, Neil Clarke5,9 , Frederic Bard6,9,#, Huck-Hui Ng1,2,3,4,9,#

1 - Gene Regulation Laboratory, Genome Institute of Singapore.
2 - School of Biological Sciences, Nanyang Technological University.
3 - Graduate School for Integrative Sciences & Engineering, National University of Singapore.
4 - Dept of Biological Sciences, National University of Singapore.
5 - Computational and Systems Biology group, Genome Institute of Singapore.
6 - Institute of Molecular and Cell Biology.
7 - Stem Cell and Developmental Biology, Genome Institute of Singapore.
8 - Center for Life Sciences, Harvard Medical School, 330 Brookline Ave, Boston, MA02115, USA.
9 - Dept of Biochemistry, National University of Singapore.
* These authors contributed equally to this work.

# Corresponding authors

Published in Nature Oct 17th, Epub ahead of print.

Abstract
The derivation of human ES cells (hESCs) from human blastocysts represents one of the milestones in stem cell biology1. The full potential of hESCs in research and clinical applications requires a detailed understanding of the genetic network that governs the unique properties of hESCs. Here, we report a genome-wide RNA interference screen to identify genes which regulate self-renewal and pluripotency properties in hESCs. Interestingly, functionally distinct complexes involved in transcriptional regulation and chromatin remodeling are among the factors identified in the screen. To understand the roles of these potential regulators of hESCs, we studied a transcription factor PRDM14 to gain new insights into its functional roles in the regulation of pluripotency. We showed that PRDM14 directly regulates the expression of key pluripotency gene POU5F1 through its proximal enhancer. Genome-wide location profiling experiments revealed that PRDM14 co-localized extensively with other key transcription factors such as OCT4, NANOG and SOX2, indicating that PRDM14 is integrated into the core transcriptional regulatory network. More importantly, in a gain-of-function assay, we showed that PRDM14 is able to enhance the efficiency of reprogramming of human fibroblasts in conjunction with OCT4, SOX2 and KLF4. Altogether, our study uncovers a wealth of novel hESC regulators wherein PRDM14 exemplifies a key transcription factor required for the maintenance of hESC identity and the reacquisition of pluripotency in human somatic cells.

 
 

 
 


Figure Legend: Schematic diagram of the experimental strategy used to identify novel regulators required to maintain human embryonic stem cell identity. a, A reporter human embryonic stem cell-line was generated for whole genome siRNA screen. b, The top candidates from the primary screen was validated. c, Informatics analyses were carried out to reveal pathways and complexes identified in the primary screen. d, Candidates required for the maintenance of hESC identity were selected for gain-of-function assay for induction of pluripotency. e, One of the candidates, PRDM14 was chosen for further mechanistic studies by mapping the binding sites on genomic DNA using ChIP-seq. f, The role of PRDM14 on transcription was analyzed.

For more information on Frederic BARD’s Lab, please click here.