ANIMAL MODELS BASED ON NUCLEAR TRANSFER IN THE RAT

描述(改编自申请人的摘要)：基因操作 建立疾病的动物模型是研究疾病的关键 人类疾病的生物学方面，是发展人类疾病所必需的 疾病的治疗干预和对自然的理解 人类多种疾病的历史。全能的发现 来自正常胚胎的胚胎干细胞是一项重要的进展，它 第一次允许人类的基因发生突变 选择。通过将同源重组与这些 细胞形成生殖系镶嵌嵌合体，已经有可能使无效 选择基因的突变，从而创建重要的动物模型 对基因在一系列生物过程中的功能的分析 人类疾病。虽然已经从小鼠的囊胚中分离出ES细胞 几种哺乳动物物种，开发有针对性的突变只在 到目前为止的鼠标。这有很多可能的原因，但 非小鼠物种在遗传学研究中的重要性，特别是大鼠， 是不可否认的。在老鼠身上完成靶向突变已被证明是 这是有问题的，因为虽然分离ES细胞是可能的，但细胞不是 能够产生生殖系镶嵌嵌合体。即使胚胎干细胞来自 大鼠囊胚可以培养并显示早期胚胎的标志 到目前为止，它们还不能用来产生遗传动物模型。 最近，核移植被证明可以从 几种哺乳动物的孤立的卵裂球。培养的细胞包括 从成年绵羊身上分离出来的那些已经被证明是可行的 当融合到去核卵子时的后代。申请者已经开发出 稳定分离大鼠早期胚胎细胞的方法 适合于体外遗传操作。申请者正在提议 利用这些细胞将核移植到受体大鼠的卵子中。这个 重建的卵子将拥有培养细胞的遗传信息， 当转移到老鼠代孕母亲的生殖道时 发育成供体核型的后代，并发生生育变化 在培养的细胞中诱导。申请人建议将这些牢房用作 一种利用快速扩展的老鼠基因组信息的工具 目的：建立新型的大鼠人类疾病动物模型。这个 在大鼠身上的大量数据使其成为许多人选择的实验模型 致癌研究、心血管研究和神经行为研究。

DESCRIPTION (Adapted from the applicant's abstract): Genetic manipulation of animals to produce animal models of disease is critical to the study of biological aspects of human disease, and is necessary for the development of therapeutic intervention in disease and an understanding of the natural history of many types of human ailment. The discovery of pluripotent embryonic stem (ES) cells from normal embryos was an important advance which allowed, for the first time, the development of mutations in genes of choice. By combining homologous recombination with the ability of these cells to form germline mosaic chimeras, it has been possible to make null mutations in genes of choice and thereby create important animal models for the analysis of the functions of genes in a range of biological process and human disease. Although ES cells have been isolated from blastocysts of several mammalian species, developing targeted mutations has only worked in the mouse to date. There are many possible reasons for this, but the importance of non-mouse species in genetic research, particularly the rat, is undeniable. Accomplishing targeted mutations in the rat has proven to be problematic, for while isolating ES cells is possible, the cells are not capable of generating germline mosaic chimeras. Even though ES cells from the rat blastocyst can be cultured and display markers of early embryonic stages, they cannot yet be used to generate genetic animal models. Recently, nuclear transfer has been shown to produce viable offspring from isolated blastomeres of several mammalian species. Cultured cells including those isolated from adult sheep have been proven to develop into viable offspring when fused to enucleated eggs. The applicants have developed methods for the stable isolation of early embryonic cells from the rat, suitable for in vitro genetic manipulation. The applicants are proposing to use these cells for nuclear transfer into recipient rat eggs. The reconstructed eggs will have the genetic information of the cultured cells, and when transferred to the reproductive tract of rat surrogate mothers will develop into offspring of the donor nuclear type, and bearing changes induced in the cultured cells. The applicants propose using these cells as a tool to take advantage of rapidly expanding rat genome information in order to establish novel animal models of human disease in the rat. The extensive data in rat make it the experimental model of choice for many carcinogenesis studies, cardiovascular studies, and neurobehavioral studies.