RAT RESOURCE AND RESEARCH CENTER: CLONING TECHNOLOGY

大鼠资源与研究中心：克隆技术

• 批准号: 7391988
• 负责人: Philip M Iannaccone
• 金额: $ 20.07万
• 依托单位: UNIVERSITY OF MISSOURI-COLUMBIA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-22 至 2007-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7391988
• 关键词: birth; cow; egg /ovum; embryonic stem cell; fibroblasts; genes; genetics; mutant; nuclear transfer; sheep; tissue /cell culture; tissue mosaicism

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The use of embryonic stem cells to gain access to the germline of experimental animals has been important to our greater understanding of gene function. As valuable as this approach is, it has not worked for species other than mouse. While ES cells have been isolated from a variety of species including the rat, the cells have not so far proven to be sufficiently pluripotent to allow the creation of germline chimeras from them. Another approach to the germline, though, is nuclear transfer. Recent advances in the use of cultured cells as a source of genetic material to direct embryonic development of enucleated oocytes in rabbit, cow, and sheep offer an approach to homologous recombination in the rat. We have established procedures for the enucleation of rat oocytes, the injection of cumulus cells and of genetically marked embryonic fibroblasts from the rat resulting in advanced preimplantation development. If it proves feasible to recover live births from this approach, then knocking-out genes in the fibroblasts can be used to obtain mutant rats. Nuclear transfer results in a rat which is essentially the strain of origin of the cultured cells without subsequent breeding and therefore allows, in principle, the use of particular strains of rat for which given experiments make the most sense.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子弹和调查员（PI）可能已经从其他NIH来源获得了主要资金，因此可以在其他清晰的条目中代表。列出的机构适用于该中心，这不一定是调查员的机构。使用胚胎干细胞进入实验动物的种系对我们对基因功能的更深刻理解很重要。尽管这种方法很有价值，但它尚未适用于小鼠以外的物种。尽管已经从包括大鼠在内的多种物种中分离出ES细胞，但迄今为止，这些细胞还没有被证明足以使它们从中产生种系嵌合体。但是，该种系的另一种方法是核转移。使用培养细胞作为遗传物质来源的最新进展，直接导致兔子，牛和绵羊中含核细胞的胚胎发育，为大鼠提供了同源重组的方法。我们已经建立了大鼠卵母细胞征核，注射积云细胞和遗传标记的胚胎成纤维细胞的方法，从而导致了晚期植入前的发育。如果事实证明可以从这种方法中恢复活产，那么可以使用成纤维细胞中的敲除基因来获得突变大鼠。核转移导致大鼠基本上是培养细胞的起源菌株，而无需随后的繁殖，因此，原则上允许使用特定的大鼠菌株对实验最有意义。