Somatic cell reprogramming by nuclear transplantation

通过核移植进行体细胞重编程

• 批准号: 262085-2011
• 负责人: Bordignon, Vilceu
• 金额: $ 2.19万
• 依托单位: McGill University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2014
• 资助国家: 加拿大
• 起止时间: 2014-01-01 至 2015-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=547982
• 关键词: Somatic; cell; reprogramming; nuclear; transplantation

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).

体细胞核移植（SCNT）到去核卵母细胞中在研究不同细胞过程的研究中具有突出的应用，以及克隆动物的实际应用（例如，拯救濒危物种、复制遗传上上级的动物、生产转基因动物、为医学研究创造动物模型）。尽管已经从体细胞克隆了20多种不同的动物，但效率仍然很低，而且仍然没有很好地理解是什么决定了SCNT产生的胚胎的成功发育或失败。由于许多原因，包括与人类的解剖学，生理学和代谢相似性，人们对应用SCNT生产转基因猪感兴趣，这些转基因猪将用作生物医学研究中的模型以研究各种条件。因此，更好地了解限制克隆成功的原因，然后利用这些研究获得的信息来建立提高克隆效率的新策略是很重要的。该研究计划的主要重点是在生产完全可行的SCNT胚胎时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损伤对体外生产胚胎的潜在影响提供新的见解，这将有助于确定体外胚胎生产的更好条件和更安全的方案。这是相关的，因为体外胚胎生产不仅广泛用于动物，而且广泛用于人类（例如，2010年诺贝尔奖授予体外受精研究）。我们提出的研究将为更可靠地选择核供体细胞（以提高克隆效率），筛选最小化DNA损伤的培养条件（以在体外产生更健康的胚胎），以及定义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）提供新的有价值的标记。