Somatic cell reprogramming by nuclear transplantation

将体细胞核移植到去核卵母细胞中，在研究不同细胞过程的研究以及克隆动物的实际应用(例如，拯救濒危物种、复制遗传优势动物、生产转基因动物、建立医学研究的动物模型)方面有着突出的应用。尽管已经从体细胞克隆了20多种不同的动物，但克隆效率仍然很低，目前还不清楚是什么决定了SCNT创造的胚胎发育成功或失败。由于许多原因，包括解剖、生理和代谢方面与人类相似，人们有兴趣将SCNT用于生产转基因猪，作为生物医学研究的模型，以研究各种条件。因此，重要的是更好地了解限制克隆成功的原因，然后利用从这些研究中获得的信息来建立提高克隆效率的新策略。这项研究计划的主要重点是在生产完全可存活的SCNT胚胎的过程中，保持移植基因组和受体宿主细胞的功能完整性。我们的研究将为DNA损伤对体外胚胎产生的潜在影响提供新的见解，这将有助于确定更好的体外胚胎生产条件和更安全的方案。这是相关的，因为体外胚胎生产不仅在动物身上得到了广泛的应用，而且在人类身上也得到了广泛的应用(例如，2010年诺贝尔奖被授予体外受精研究)。我们的研究将为更可靠地选择核供体细胞(提高克隆效率)提供新的有价值的标记，为筛选将DNA损伤降至最低的培养条件(以在体外产生更健康的胚胎)提供新的有价值的标记，并为确定新的和改进的策略来增强染色质重编程(以提高从SCNT克隆动物的效率)。

Somatic cell nuclear transfer (SCNT) into enucleated oocytes has outstanding applications in research to study different cellular processes, as well as practical applications for cloning animals (e.g., save endangered species, replicate genetically superior animals, produce genetically modified animals, create animal models for medical research). Although more than 20 different species of animals have been cloned from somatic cells, the efficiency remains low and it is still not well understood what determines either successful development or failure in embryos created by SCNT. Because of a number of reasons that includes anatomical, physiological and metabolic similarities with humans, there is interest in the application of SCNT for the production of transgenic pigs to be used as models in biomedical research to study a variety of conditions. Therefore, it is important to gain a better understanding of the causes that limit the cloning success and then use the information obtained from these studies to establish novel strategies for increasing cloning efficiency. The major focus of this research program is on the preservation of the functional integrity of both the transplanted genome and recipient host cell in the production of fully viable SCNT embryos. Our studies will provide new insights on the potential effect of DNA damage on embryos produced in vitro, which will help on defining better conditions and safer protocols for in vitro embryo production. This is relevant because in vitro embryo production is used extensively not only in animals and but also in humans (e.g., the 2010 Nobel Prize was awarded for studies on in vitro fertilization). Our proposed studies will provide new valuable markers for more reliable selection of nuclear donor cells (to increase the efficiency of cloning), for the screening of culture conditions that minimize DNA damage (to produce healthier embryos in vitro), and for defining new and improved strategies to enhance chromatin reprogramming (to increase the efficiency of animal cloning from SCNT).