Utilizing single cell approaches to decipher the mechanism underlying somatic cel

描述（由申请人提供）：了解表征体细胞经历重编程为多能性的基本分子元件是将干细胞研究推向临床应用的先决条件。该领域的一个重要问题是，重编程过程是否是由随机事件驱动的，这些事件没有特定的序列发生，或者它是一个更具确定性的过程，具有定义的转录变化模式，必须顺序发生，以将分化的细胞转化为诱导多能干细胞（iPSC）。破译这些元素的主要障碍之一是转导细胞群的异质性和最终将产生iPSC的细胞的小部分。事实上，该领域的大多数研究都是基于细胞群分析，这可能并不代表经历重编程的细胞的一小部分，这强烈表明，利用单细胞技术对于理解重编程过程至关重要。因此，我的博士后培训的主要目标将是使用一种创新的单细胞技术来破译定义重编程过程的分子机制，该技术可以监测单细胞的转录谱。这项技术将使我了解在重编程过程中发生的主要事件是什么，它们发生的时间顺序是什么，以及哪些基因负责执行这些事件。然后，我将描述事件和关键参与者，这些事件和关键参与者将在我的分析中突出显示，以了解他们如何在重编程过程中执行其功能。最后，我将使用突出显示的基因生成敲入二级系统，用于未来的重编程研究。解决这些问题将有助于阐明重编程过程的分子机制，并将成为确定重编程过程是确定性还是随机性的有力工具。

DESCRIPTION (provided by applicant): Understanding the basic molecular elements that characterize somatic cells undergoing reprogramming to pluripotency is a prerequisite for advancing stem cell research towards clinical applications. One of the important questions in the field is whether the reprogramming process is driven by stochastic events that occur in no particular sequence or rather it is a more deterministic process that has a defined pattern of transcriptional changes that must occur sequentially to convert the differentiated cells to induced pluripotent stem cells (iPSCs). One of the major obstacles in deciphering these elements is the heterogeneity of the transduced cell population and the small fraction of cells that eventually will yield iPSCs. The fact that the majority of the studies in the field are based on cell-population analyses, which presumably do not represent the small fraction of cells that undergo reprogramming, strongly suggests that utilizing single cell techniques will be of paramount importance to understand the reprogramming process. Therefore, the main objective of my postdoctoral training will be to decipher the molecular mechanisms that define the reprogramming process using an innovative single cell technique that monitors the transcriptional profile of single cells. This technique will allow me to understand what the main events that occur during the reprogramming process are, in what chronological order they occur, and which genes are responsible for executing these events. I will then characterize the events and the key players that will be highlighted in my analysis to understand how they execute their function in the reprogramming process. Finally, I will generate knock-in secondary systems using the highlighted genes for future reprogramming studies. Addressing these questions will shed light on the molecular mechanisms that underlie the reprogramming process and will be a powerful tool to determine whether the reprogramming process is deterministic or stochastic.