TransChromo Mouse™
TransChromo Mouse™
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转色鼠标™
DOI：
10.1080/02648725.2002.10648023
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发表时间：
2002
影响因子：
--
通讯作者：
Yoshimi Kuroiwa
中科院分区：
文献类型：
--
作者：
Isao Ishida;Kazuma Tomizuka;Hitoshi Yoshida;Yoshimi Kuroiwa
利用基因操纵小鼠胚胎干细胞产生嵌合体的各种技术为研究基因在体内的功能和基因表达的发育调控提供了有价值的工具。通过同源重组对特定基因进行靶向修饰（Mansour et al., 1988），引入克隆的外源DNA片段（Jakobovits et al., 1993; Strauss et al., 1993），以及通过Cre-loxP系统对染色体进行修饰（ramez - solis el aI）。, 1995;史密斯和我。（1995）为胚胎干细胞的基因操纵提供了便利。细胞融合或染色体转移是表征体细胞中基因表达的不可或缺的工具（Hooper, 1987）。虽然已经报道了一些使用胚胎癌（Ee）细胞的此类研究（Benham et al., 1983），但所描述的技术很少用于操纵胚胎干（ES）细胞。利用转染色体胚胎干细胞培育出具有特定外源染色体介导功能的动物，使得在体内研究非常大的基因或基因簇成为可能，这些基因或基因簇远远超过了传统技术所能克隆的DNA的有限长度。由于它们的大尺寸，这些HACs允许包含关键的relnote调节元件，从而使我们能够重现适当表达转荧光元件所需的条件。这种方法为构建涉及氯硝索酮异常的人类遗传疾病的动物模型提供了有吸引力的可能性。20多年前，Ilhnensee及其同事报道了将杂交小鼠EC细胞与人纤维肉瘤或大鼠肝细胞融合产生可存活的儿童小鼠（I1hllensee et al.•1978;Ilhnensee and Croce, 1979）。然而,他们
Various techniques for producing chiJneras froIII genetically manipulated mouse ES cells have provided valuable tools for studying gene functions ill vivo and developmental regulation of gene expression. A variety of procedures, such as targetted Illodification of specific genes by homologous recombination (Mansour et al., 1988), introduction of cloned exogenous DNA fraglnents (Jakobovits et al., 1.993; Strauss et al., 1993) and chromosonle nlanipu)ation by the Cre-loxP system (Ramiez-Solis el aI., 1995; Smith et aI., 1995) have facilitated the genetic Illanipulation of ES cells. Cell fusion, or chrOlnosome transfer, is an indispensable tool for characterizing gene expression in somatic cells (Hooper, 1987). Although several such studies using elnbryonic carcinoma (Ee) cells (Benham et al., 1983) have been reported, the techniques described have rarely been used to manipulate embryonic stem (ES) cells. The creation of animals Inanifesting specific foreign chromosolne-Inediated functions using transchromosolnic ES cells allows the study in vivo of very large genes or gene clusters that far exceed the limited length of DNA that can be cloned by conventional techniques. Because of their large size t these HACs allow the inclusion of critical relnote regulatory eleillents and thereby pennit us to reproduce the conditions necessary for the proper expression of the transchroJnosolnic elements. This approach opens attractive possibilities forconstrucling aniInallnodels representing human genetic diseases involving chrolnosolne abnonnalities. More than 20 years ago, Ilhnensee and co-workers reporled the production of viable chilneric mice froln hybrid rnouse EC cells fused with human fibrosarcoma or rat hepatollla cells (I1hllensee et al.• 1978; Ilhnensee and Croce, 1979). However, they