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NATIONAL INSTITUTE OF GENETICS
Mammalian Development Laboratory

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RESEARCH

Reserch

  1. Genetic technologies in the mouse
  2. Molecular mechanisms of somite segmentation
  3. Germ cell development
  4. The roles of Notch signaling in cardiovascular development

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1. Genetic technologies in the mouse

Gene targeting

To understand the functions of genes expressed during development, we employ several methods for generating genetically modified mice. One of the principal techniques that we utilize is Embryonic Stem (ES) cell mediated gene targeting (Figs. 1 & 2).

This gene targeting strategy is based on two critical events: 1) homologous recombination in ES cells and 2) germline transmission through the chimeric mouse. For generating chimeric mice, the two main methods are aggregation, in which targeted ES cells are co-cultured with 8 cell-stage host embryos, and microinjection, in which ES cells are injected directly into the blastocyst of the host embryo.

We find the aggregation method preferable, as it does not require high levels of technical expertise, and using this protocol we have successfully generated both gene-knockout and -knockin mice.

Figure1: Establishment of a gene targeted ES cell line

Figure2:Generation of chimera mice using aggregation method

Bac modified transgenic technologies

Transgenic technologies are also, however, important and powerful methods for generating genetically modified mice. Recently we have developed an efficient BAC modification method, by which we can integrate any gene into a desired locus using a bacterial homologous recombination system (Fig. 3). This system can be used successfully in gene expression analyses and in studies that examine the global regulation of transcription.

ノックインマウスの作出

これは基本的にはノックアウトマウスの作成と全く同じ原理を利用する。遺伝子を破壊するかわりに別の遺伝子を任意に導入することが可能である。たとえば遺伝子の発現をモニターするために LacZ 遺伝子や GFP を導入することができる。また遺伝子に変異を導入して入れ替えることも可能である。

 

これらの方法を用いて、多くの疾患モデルマウスが作出されている。






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