ANIMAL MODELS BASED ON NUCLEAR TRANSFER IN THE RAT

描述（改编自申请人摘要）：基因操作 动物的动物模型，以产生疾病的动物模型是至关重要的研究， 人类疾病的生物学方面，并且是发展所必需的。 疾病的治疗干预和对自然的理解 许多人类疾病的历史。 多能性的发现 来自正常胚胎的胚胎干细胞是一个重要的进展， 第一次允许了基因突变的发展， 选择 通过将同源重组与这些 细胞形成生殖系嵌合体，已经有可能使无效 选择的基因突变，从而创造了重要的动物模型， 分析基因在一系列生物过程中的功能， 人类疾病 虽然胚胎干细胞已经从胚胎干细胞的囊胚中分离出来， 几个哺乳动物物种，发展有针对性的突变只在 老鼠到目前为止 有很多可能的原因，但 非小鼠物种在遗传研究中的重要性，特别是大鼠， 不可否认 在大鼠中完成靶向突变已被证明是 问题是，虽然分离ES细胞是可能的，但细胞不是 能够产生生殖系嵌合体。 尽管胚胎干细胞 大鼠囊胚可培养并显示早期胚胎标记 阶段，它们还不能用于产生遗传动物模型。 最近，核移植已被证明可以产生可存活的后代 几种哺乳动物的分离的卵裂球。 培养的细胞包括 那些从成年绵羊中分离出来的已经被证明可以发展成 与去核卵融合的后代。 申请人已经开发了 从大鼠中稳定分离早期胚胎细胞的方法， 适合于体外遗传操作。 申请人建议 将这些细胞核移植到受体大鼠卵细胞中。 的 重建的卵子将具有培养细胞的遗传信息， 当转移到大鼠代孕母亲的生殖道时， 发育成供体细胞核类型的后代， 在培养的细胞中诱导。 申请人建议使用这些细胞作为 利用快速扩展的大鼠基因组信息， 建立新型的人类疾病大鼠动物模型。 的 在大鼠中的广泛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.