DNA repair pathway choice mediates somatic cell reprogramming

项目摘要 重编程是将体细胞转化为多能干细胞。这项技术现在已经成为常规 在世界各地的实验室中使用，但绝大多数重编程干细胞系不是 发育完全胜任，损害他们在研究和治疗中的效用。重编程诱导 DNA损伤，这可能对所产生的细胞的质量产生持久的影响。我们的研究 表明重编程过程中的DNA损伤是由DNA复制异常引起的。但 DNA损伤的原因，修复的机制，以及损伤的发育后果， 没有被很好地理解。这个建议的优点是，使用实验系统，我们能够 识别由重编程诱导的特定类型的损伤，以及所需的分子机制， 修复：我们能够区分双链断裂HR的作用和停滞的复制叉的作用。 稳定我们还能够区分基因组不稳定性对重编程效率的影响， 不完全的转录转换。我们能够绘制基因组中的位点， 我们能够识别可用于增加基因组稳定性的途径， 提高重编程干细胞发育能力。这些研究将提供一个机制 了解基因组不稳定性如何抑制不同细胞状态之间的诱导转换。

Project Summary Reprogramming is the conversion of a somatic cell to a pluripotent stem cell. This technique is now routinely used in laboratories around the world, but the vast majority of reprogrammed stem cell lines are not developmentally fully competent, compromising their utility in research and therapy. Reprogramming induces DNA damage, which can have lasting consequences on the quality of the resulting cells. Our studies have shown that DNA damage during reprogramming is induced by abnormalities in DNA replication. However, the cause of DNA damage, the mechanisms of repair, and the developmental consequences of the damage are not well understood. The strength of this proposal is that with the experimental systems used, we are able to identify the specific type of damage induced by reprogramming, and the molecular mechanisms required for repair: we are able to distinguish the role of double strand break HR from the role of stalled replication fork stability. We are also able to distinguish the effect of genome instability on reprogramming efficiency from incomplete transcriptional transitions. We are able to map the sites in the genome with reprogramming-induced damage, and we are able to identify pathways that can be used to increase genome stability and potentially improve developmental competence of reprogrammed stem cells. These studies will provide a mechanistic understanding how genome instability inhibits the induced transition between different cellular states.