Analysis of Mitotic and Expression Properties of Human Neocentromere-based Transchromosomes in Mice*
Analysis of Mitotic and Expression Properties of Human Neocentromere-based Transchromosomes in Mice*
复制标题
小鼠基于人新着丝粒的转染色体的有丝分裂和表达特性分析*
DOI：
--
复制
发表时间：
2005
影响因子：
4.8
通讯作者：
K. Choo
中科院分区：
文献类型：
--
作者：
L. Wong;R. Saffery;M. Anderson;E. Earle;Julie M. Quach;A. Stafford;K. Fowler;K. Choo
人类新着丝粒是一种功能性着丝粒，不含典型的人类着丝粒α-卫星DNA。我们已经将60-Mb的10号染色体衍生的新着丝粒标记染色体martel（10）及其截短的3.5-Mb衍生物NC-MiC 1转移到小鼠胚胎干细胞中，并证明了在异源遗传背景下转染色体的相对高的结构和有丝分裂稳定性。我们还生产了携带martel（10）或NC-MiC 1的嵌合小鼠。两种转染色体在包括造血干细胞在内的多种成年嵌合小鼠组织中均被检测为完整的附加体实体。位于这些转染色体上的基因在测试的不同组织中表达。从精子样本中这些染色体的DNA检测中可以看出，嵌合小鼠中两种转染色体的减数分裂传递是明显的。特别是，在嵌合小鼠的F1胚胎中证明了NC-MiC 1的生殖系传播。检测到这些转染色体的错误分离水平可变（在含martel（10）或NC-MiC 1的胚胎干细胞和嵌合小鼠组织中较低，而在含NC-MiC 1的F1胚胎中相对较高），表明它们在小鼠背景中并非最佳倾向于完全有丝分裂调控，特别是在早期胚胎发生期间。这些结果提供了有前途的数据，支持潜在的使用基于新着丝粒的人类标记染色体和微型染色体作为工具的着丝粒，新着丝粒和染色体生物学的研究，并在小鼠模型系统的基因治疗研究。他们还强调了需要进一步了解和克服的因素，负责这些转染色体在小鼠模型中的可定义的不稳定率。
Human neocentromeres are functional centromeres that are devoid of the typical human centromeric α-satellite DNA. We have transferred a 60-Mb chromosome 10-derived neocentric marker chromosome, mardel(10), and its truncated 3.5-Mb derivative, NC-MiC1, into mouse embryonic stem cell and have demonstrated a relatively high structural and mitotic stability of the transchromosomes in a heterologous genetic background. We have also produced chimeric mice carrying mardel(10) or NC-MiC1. Both transchromosomes were detected as intact episomal entities in a variety of adult chimeric mouse tissues including hemopoietic stem cells. Genes residing on these transchromosomes were expressed in the different tissues tested. Meiotic transmission of both transchromosomes in the chimeric mice was evident from the detection of DNA from these chromosomes in sperm samples. In particular, germ line transmission of NC-MiC1 was demonstrated in the F1 embryos of the chimeric mice. Variable (low in mardel(10)- or NC-MiC1-containing embryonic stem cells and chimeric mouse tissues and relatively high in NC-MiC1-containing F1 embryos) levels of missegregation of these transchromosomes were detected, suggesting that they are not optimally predisposed to full mitotic regulation in the mouse background, particularly during early embryogenesis. These results provide promising data in support of the potential use of neocentromere-based human marker chromosomes and minichromosomes as a tool for the study of centromere, neocentromere, and chromosome biology and for gene therapy studies in a mouse model system. They also highlight the need to further understand and overcome the factors that are responsible for the definable rates of instability of these transchromosomes in a mouse model.