A genome-wide RNAi screen reveals determinants of human embryonic stem cell identity.
The derivation of human ES cells (hESCs) from human blastocysts represents one of the milestones in stem cell biology. 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 remodelling are among the factors identified in the screen. To understand the roles of these potential regulators of hESCs, we studied transcription factor PRDM14 to gain new insights into its functional roles in the regulation of pluripotency. We showed that PRDM14 regulates directly the expression of key pluripotency gene POU5F1 through its proximal enhancer. Genome-wide location profiling experiments revealed that PRDM14 colocalized 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.
Chia
Na-Yu
NY
Gene Regulation Laboratory, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672.
Chan
Yun-Shen
YS
Feng
Bo
B
Lu
Xinyi
X
Orlov
Yuriy L
YL
Moreau
Dimitri
D
Kumar
Pankaj
P
Yang
Lin
L
Jiang
Jianming
J
Lau
Mei-Sheng
MS
Huss
Mikael
M
Soh
Boon-Seng
BS
Kraus
Petra
P
Li
Pin
P
Lufkin
Thomas
T
Lim
Bing
B
Clarke
Neil D
ND
Bard
Frederic
F
Ng
Huck-Hui
HH
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D065150:Cellular Reprogramming / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D004742:Enhancer Elements, Genetic / Q000235:genetics; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005786:Gene Expression Regulation / Q000235:genetics; D015894:Genome, Human / Q000235:genetics*; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D000090062:Kruppel-Like Factor 4; D051379:Mice; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D016601:RNA-Binding Proteins; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D014157:Transcription Factors
D004268:DNA-Binding Proteins; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; C072725:NFRKB protein, human; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; C524724:PRDM14 protein, human; D016601:RNA-Binding Proteins; D012097:Repressor Proteins; C527782:SOX2 protein, human; D055748:SOXB1 Transcription Factors; D014157:Transcription Factors