Journal Article

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

Lu, Xinyi, Bard, Frederic, Feng, Bo, Li, Pin, Orlov, Yuriy L., Lau, Mei-Sheng, Huss, Mikael, Kraus, Petra, Jiang, Jianming, Lim, Bing, Moreau, Dimitri, Yang, Lin, Lufkin, Thomas, Kumar, Pankaj, Clarke, Neil D., Ng, Huck-Hui, Chia, Na-Yu, Soh, Boon-Seng, Chan, Yun-Shen

2010

Overview

Screen test for stem-cell cues If the full potential of human embryonic stem (ES) cells in research and clinical applications is to be realized, we need a detailed understanding of the genetic network that governs their unique properties. A genome-wide RNA interference screen identifies a wealth of regulators of self-renewal and pluripotency properties in human embryonic stem cells. PRDM14, for example, represents a key transcription factor required for the maintenance of human ES cell identity and reprogramming of somatic cells to pluripotency. Realizing the full potential of human embryonic stem cells (hESCs) in research and clinical applications requires a detailed understanding of the genetic network that governs their unique properties. A genome-wide RNA interference screen identifies a wealth of new regulators of self-renewal and pluripotency properties in hESCs. The transcription factor PRDM14, for example, is required for the maintenance of hESC identity and reprogramming of somatic cells to pluripotency. The derivation of human ES cells (hESCs) from human blastocysts represents one of the milestones in stem cell biology 1 . 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.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 631/208/191/505 / 631/532/2064/2117 / 692/308/2171 / Animals / Base Sequence / Biological and medical sciences / Candidates / Cell differentiation, maturation, development, hematopoiesis / Cell Line / Cell physiology / Cellular Reprogramming - genetics / DNA-Binding Proteins - genetics / DNA-Binding Proteins - metabolism / Embryonic stem cells / Embryonic Stem Cells - cytology / Embryonic Stem Cells - metabolism / Enhancer Elements, Genetic - genetics / Fibroblasts - cytology / Fibroblasts - metabolism / Fundamental and applied biological sciences. Psychology / Gene expression / Gene Expression Regulation - genetics / Genome, Human - genetics / Humanities and Social Sciences / Humans / Induced Pluripotent Stem Cells - cytology / Induced Pluripotent Stem Cells - metabolism / letter / Mice / Molecular and cellular biology / multidisciplinary / Octamer Transcription Factor-3 - genetics / Octamer Transcription Factor-3 - metabolism / Physiological aspects / Proteins / Repressor Proteins - genetics / Repressor Proteins - metabolism / RNA Interference / Science / Science (multidisciplinary) / SOXB1 Transcription Factors - metabolism / Stem cell research / Stem cells