Identifying regulators of the naive and primed pluripotent state

识别幼稚和启动多能状态的调节因子

• 批准号: 8784669
• 负责人: Inna Lipchina
• 金额: $ 5.33万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2017-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8784669
• 关键词: Address; Biochemical; Biological Assay; Cell Line; Cell Transplantation; Cell surface; Cells; DNA Methylation; Derivation procedure; Development; Disease model; Doxycycline; Embryo; Epiblast; Epigenetic Process; Event; Gene Targeting; Generations; Human; In Vitro; Individual; Investigation; Libraries; Maintenance; Mus; Pathway interactions; Population; Preclinical Drug Evaluation; Process; Refractory; Regulation; Reporter; Research; Signal Transduction; Staging; Stem cells; System; Testing; Transgenic Organisms; blastocyst; cell type; embryonic stem cell; gene discovery; genome wide association study; genome-wide; histone modification; human embryonic stem cell; implantation; novel; overexpression; pluripotency; preimplantation; public health relevance; self-renewal; small hairpin RNA; tool; transcription factor; vector

DESCRIPTION (provided by applicant): Recent evidence suggests that mouse embryonic stem cells can self-renew in different states of pluripotency, embryonic stem cells (ESCs) and epiblast stem cells (EpiSCs), which developmentally correspond to pre- and post-implantation stage epiblast of the embryo, respectively. Exogenous expression of individual transcription factors has been shown to be sufficient to reprogram EpiSCs into na¿ve ESCs. However, despite the developmental similarities of these pluripotent cell types, conversion efficiencies remain low and little is known about the underlying transcriptional, epigenetic, and signaling events of this process. In order to elucidate the mechanisms of EpiSC-to-ESC reprogramming, we here propose to (i) establish fluorescent reporter and inducible transgenic tools to effectively study this process, (ii) define intermediate stages of EpiSC-to-ESC conversion at the transcriptional and epigenetic level, and (iii) perform an unbiased genome-wide shRNA screen to identify novel roadblocks of this process. The investigation of the mechanism of EpiSC-to-ESC conversion is a powerful platform from which we can discover genes important during pre-implantation development and the acquisition of pluripotency. Moreover, we expect that our studies will provide important clues for stabilizing a na¿ve pluripotent state in human cells.

描述（由申请人提供）：最近的证据表明，小鼠胚胎干细胞可以在不同的多能状态下自我更新，胚胎干细胞（ESCs）和外胚层干细胞（EpiSCs），它们在发育上分别对应于胚胎着床前和着床后的外胚层。单个转录因子的外源表达已被证明足以将EpiSCs重编程为na - ve ESCs。然而，尽管这些多能细胞类型的发育相似，但转换效率仍然很低，并且对这一过程的潜在转录、表观遗传和信号事件知之甚少。为了阐明episc_to - esc重编程的机制，我们提出(i)建立荧光报告和可诱导的转基因工具来有效地表达episc_to - esc重编程